1.

A sample of human DNA is subjected to increasing temperature until the major fraction
exhibits optical density changes due to disruption of its helix (melting or denaturation). A
smaller fraction is atypical in that it requires a much higher temperature for melting. This
smaller atypical fraction of DNA must contain a higher content of
A. Adenine plus cytosine
!. "ytosine plus guanine
". Adenine plus thymine
D. "ytosine plus thymine
#. Adenine plus guanine
The answer is: B
The melting temperature Tm of duplex DNA is the temperature at $hich half the base
pairs are denatured. Adenine%thymine (A%T) base pairs ha&e t$o hydrogen bonds in
contrast to cytosine%guanine ("%') base pairs $hich ha&e three hydrogen bonds. Duplex
DNA molecules rich in A%T base pairs ha&e a much lo$er Tm than those rich in "%' base
pairs. As DNA is heated fractions $ith a higher A%T content melt or denature before
those $ith a higher "%' content. (ost mammals including humans ha&e satellite DNA
fractions that are highly repetiti&e and clustered in particular chromosome regions.
)atellite DNAs are named for their altered density (satellite band) on centrifugation
caused by higher '%" content. Their function is un*no$n.
+.
A ne$born baby has a sibling $ith sic*le cell anemia (1,1-..) and is at ris* for the
disease. /hich of the follo$ing is the appropriate diagnostic test for sic*le cell anemia in
this baby0
A. DNA amplification
!. 1emoglobin antibodies
". DNA restriction
D. 2ed cell counting
#. DNA fingerprinting
The answer is: A
)ufficient DNA for analysis can be obtained by amplification of leu*ocyte DNA using the
polymerase chain reaction (3"2). )hort segments of DNA (oligonucleotide primers) are
designed to be complementary to areas flan*ing the DNA region of interest4in this case
the portion of the %globin gene that may harbor the sic*le cell anemia (1,1-..)
mutation. )ome +. to 5. cycles of cooling (to anneal the primers) synthesis ($ith heat%
stable DNA polymerase) and heating (to melt the DNA and allo$ the next cycle) can
amplify the targeted DNA segment o&er 1 million%fold. 1ybridi6ation in duplicate to
allele%specific oligonucleotides (A)7s8 one A)7 for the hemoglobin A mutation one
A)7 for the ) mutation) can establish the diagnosis of normal (AA alleles) sic*le trait
(A) alleles) or sic*le cell anemia ()) alleles). Ne$borns ha&e fetal hemoglobin ( % and
%globin) $ith little expression of hemoglobin A ( % and %globin genes) until 5 to 9
months of life so testing for anemia or for abnormal hemoglobin $ith antibodies $ould
not be helpful.
DNA polymorphisms (nucleotide sequence &ariations) occur approximately once per +..
to :.. base pairs (bp) of human DNA. ;f the sequence &ariation affects the recognition
site for a restriction endonuclease the altered segment si6es produced by endonuclease
digestion allo$ detection of the sequence change <restriction fragment length
polymorphism (2=>3)?. ;f the nucleotide change causing the 2=>3 is adjacent to (lin*ed
$ith) or coincident $ith a disease mutation then one si6e &ariant of the 2=>3 may be
diagnostic. 1o$e&er mutations of *no$n sequence (such as that for sic*le cell anemia)
are better detected by 3"2 and A)7s. The use of se&eral highly &ariable 2=>3s produces
a pattern of restriction fragments that is highly distincti&e for each indi&idual (DNA
fingerprinting) but not diagnostic for a particular disease.
5. A farming couple in Northern (ichigan consult their physician about se&ere s*in
rashes and ulcers noted o&er the past year. They also ha&e lost many cattle o&er the past
year and claim that their cattle feed changed in consistency and smell about 1 year ago.
"hemical analysis of the feed sho$s high concentrations of polychlorinated biphenyls a
fertili6er related to *no$n carcinogens. The physician sends the chemical to a laboratory
for carcinogen testing $hich is performed initially and rapidly by
A. ;noculation of the chemical into nude mice
!. ;ncubation of mutant bacteria $ith the chemical to measure the rate of re&erse or
@bac*@ mutations
". ;ncubation $ith stimulated $hite blood cells to measure the impact on DNA
replication
D. "omputer modeling based on the structures of related carcinogens
#. ;ncubation $ith mammalian cell cultures to measure the rates of malignant
transformation
The answer is: B
The Ames test is a rapid and relati&ely inexpensi&e bacterial assay for determining
mutagenicity of potential toxic chemicals. )ince many chemical carcinogens are
mutagenic it seems ob&ious that damage to DNA is a central e&ent in carcinogenesis as
$ell as mutagenesis. Dr. !ruce Ames de&eloped a tester strain of Salmonella that has
been modified not to gro$ in the absence of histidine because of a mutation in one of the
genes for the biosynthesis of histidine. Toxic chemicals that are mutagens are placed in
the center of the plate and result in re&ersions of the original mutations so that histidine
is synthesi6ed and the mutated re&ertants multiply in histidine%free media. )ince many
carcinogens are con&erted to acti&e forms by metabolism in the li&er preliminary
incubation $ith li&er homogenates may precede the bacterial assay. #ssentially all
chemicals *no$n as carcinogens in humans cause mutagenesis in the Ames test. The
other options4carcinogenicity screening in immunosuppressed (nude) mice computer
modeling or incubation $ith mammalian cell cultures4may pro&ide some information
but are less efficient and &alidated than the Ames test. "ontamination of (ichigan cattle
feed $ith polychlorinated biphenyls (3"!s) did occur through an industrial mista*e.
,.
3atients $ith hereditary nonpolyposis colon cancer <1N3"" (11,:..)? ha&e genes $ith
microsatellite instability that is many regions containing abnormal small loops of
unpaired DNA. This is a result of a mutation affecting
A. (ismatch repair
!. "hain brea* repair
". !ase excision repair
D. Depurination repair
#. Nucleotide excision repair
The answer is: A
7ne of the most common types of inherited cancers is nonpolyposis colon cancer
<1N3"" (11,:..)?. (ost cases are associated $ith mutations of either of t$o genes that
encode proteins critical in the sur&eillance of mismatches. (ismatches are due to copying
errors leading to one% to fi&e%base unmatched pieces of DNA. T$o% to fi&e%base%long
unmatched bases form miniloops. Normally specific proteins sur&ey ne$ly formed DNA
bet$een adenine methylated bases $ithin a 'AT" sequence. (ismatches are remo&ed
and replaced. =irst a 'AT" endonuclease nic*s the faulty strand at a site complementary
to 'AT" then an exonuclease digests the strand from the 'AT" site beyond the
mutation. =inally the excised faulty DNA is replaced. ;n 1N3"" the unrecogni6ed
mismatches accumulate leading to malignant gro$th of colon epithelium. The other
forms of DNA repair are important for rectifying damage from ultra&iolet light.
:.
A culture of bacteria not resistant to tetracycline de&elops an infection from a &irus that is
deri&ed from the lysis of tetracycline%resistant bacteria. (ost of the bacterial progeny of
the original culture is found to ha&e become resistant to tetracycline. /hat phenomenon
has occurred0
A. "onjugation
!. "olinearity
". 2ecombination
D. Transformation
#. Transduction
The answer is: E
The process of transduction in&ol&es the transfer of a portion of DNA from one bacterium
to the chromosome of another bacterium by means of a &iral infection. "onjugation is the
transfer of a so%called male chromosomal DNA to the DNA of an acceptor or female
bacterial cell. "olinearity defines the relationship bet$een genes and proteins in that the
sequence of amino acids in proteins is a result of the sequence of base triplets in template
genes. 2ecombination is simply the exchange of sequences bet$een t$o molecules of
DNA. Transformation results $hen exogenous DNA fragments are incorporated into the
chromosome of another organism as in the transformation of pneumococcal bacteria that
led A&ery and (c>eod to recogni6e the genetic significance of DNA
9.
/hich of the follo$ing molecules is found in a nucleoside0
A. A pyrophosphate group
!. A 1A base lin*ed to a pentose sugar
". A :A%phosphate group lin*ed to a pentose sugar
D. A 5A%phosphate group lin*ed to a pentose sugar
#. A terminal triphosphate
The answer is: B
A nucleoside consists of a purine or pyrimidine base lin*ed to a pentose sugar. The 1A
carbon of the pentose is lin*ed to the nitrogen of the base. ;n DNA +A%deoxyribose sugars
are used8 in 2NA ribose sugars are used. Nucleotides are phosphate esters of nucleosides
$ith one to three phosphate groups such as adenosine monophosphate (A(3) adenosine
diphosphate (AD3) or adenosine triphosphate (AT3). The nitrogenous bases are adenine
thymine guanine and cytosine in DNA $ith thymine replaced by uridine in 2NA.
Nucleotide polymers are chains of nucleotides $ith single phosphate groups joined by
bonds bet$een the 5A%hydroxyl of the preceding pentose and the :A%phosphate of the next
pentose. 3olymeri6ation requires high%energy nucleotide triphosphate precursors that
liberate pyrophosphate (bro*en do$n to phosphate) during joining. The polymeri6ation
reaction is gi&en specificity by complementary 2NA or DNA templates and rapidity by
en6yme catalysts called polymerases.
B.
/hich of the follo$ing en6ymes can be described as a DNA%dependent 2NA
polymerase0
A. DNA ligase
!. 3rimase
". DNA polymerase ;;;
D. DNA polymerase ;
#. 2e&erse transcriptase
The answer is: B
3rimase is a DNA%dependent 2NA polymerase located in the primosome at the
replication for* of DNA. 3rimase initiates DNA synthesis by synthesi6ing a 1.%base 2NA
primer. The DNA%2NA helix formed binds DNA polymerase ;;; $hich synthesi6es a
DNA fragment (the 7*a6a*i fragment) in a :A to 5A direction. /hen the 2NA primer of
the pre&ious 7*a6a*i fragment is met DNA polymerase ; replaces ;;; and digests the
2NA primer replacing it $ith appropriate DNA bases. /hen the 2NA primer is
completely remo&ed DNA ligase synthesi6es the last phosphodiester bond thereby
sealing the space. /hat is left is a ne$ lagging strand extended by the ne$ 7*a6a*i
fragment $ith the 1.%base 2NA primer at its :A end. 2e&erse transcriptase is a DNA
polymerase that uses 2NA as a template found in retro&iruses as $ell as normal
eu*aryotic cells. Cnli*e DNA polymerase ; and ;;; $hich proofread for errors during
normal synthesis re&erse transcriptase has no proofreading capabilities. 1ence it has an
exceedingly high error rate that contributes to the high rate of mutation in retro&iruses
li*e 1;D.
E.
The DNA sequence ( sho$n belo$ is the sense strand from a coding region *no$n to
be a mutational @hot spot@ for a gene. ;t encodes amino acids +1 to +:. 'i&en the genetic
and amino acid codes """ F proline (3) '"" F alanine (A) TT" F phenylalanine (=)
and TA' F stop codon $hich of the follo$ing sequences is a frame%shift mutation that
causes termination of the encoded protein0
( :A%"""%""T%A''%TT"%A''%5A
A. %""A%""T%A''%TT"%A''%
!. %'""%""T%A''%TT"%A''%
". %""A%"""%TA'%'TT%"A'%
D. %"""%"TA%''T%T"A%''4
#. %"""%""T%A''%A''44
The answer is: C
;nsertion (choice c) or deletion (choice d) of nucleotides shifts the reading frame unless
the change is a multiple of 5 (choice e). =rame shifts may create unintended stop codons
as in choice c. 3oint mutations resulting in nucleotide or amino acid substitutions are
con&eniently named by their position in the protein i.e. 3+1A (choice b). The protein
change 3+1A could also be denoted by the corresponding change in the DNA reading
frame i.e. "95A. Deletions may be prefixed by the letter delta as $ith =+: (choice e).
-.
The hypothetical @stimulin@ gene contains t$o exons that encode a protein of 1.. amino
acids. They are separated by an intron of 1.. bp beginning after the codon for amino acid
1.. )timulin messenger 2NA (m2NA) has :A and 5A untranslated regions of B. and 5.
nucleotides respecti&ely. A complementary DNA (cDNA) made from mature stimulin
2NA $ould ha&e $hich of the follo$ing si6es0
A. :.. bp
!. ,.. bp
". 5.. bp
D. 1.. bp
#. B. bp
The ans$er isG !
#xons are the coding portions of genes and consist of trinucleotide codons that guide the
placement of specific amino acids into protein. ;ntrons are the noncoding portions of
genes that may function in e&olution to pro&ide @shuffling@ of exons to produce ne$
proteins. The primary 2NA transcript contains both exons and introns but the latter are
remo&ed by 2NA splicing. The :A (upstream) and 5A (do$nstream) untranslated 2NA
regions remain in the mature 2NA and are thought to regulate 2NA transport or
translation. A poly(A) tail is added to the primary transcript after transcription $hich
facilitates transport and processing from the nucleus. The disco&ery of introns
complicated (endelAs idea of the gene as the smallest hereditary unit8 a modern definition
might be the colinear sequence of exons introns and adjacent regulatory sequences that
accomplish protein expression. Csing these principles one can determine the si6e of the
stimulin gene. ;t contains a coding region of 5.. bp (1.. amino acids H 5 bp per amino
acid) plus 1.. bp in the intron plus B. I 5. F 1.. bp in the untranslated regions (total F
:.. bp). The mature 2NA contains the same number of bp except for the 1.. bp in the
intron (:.. J 1.. F ,.. bp). Transcription begins at the start of the :A untranslated region
(B. bp) and the splice site occurs 5. bp (1. H 5) into the coding region at the beginning of
the intron.
1..
The hypothetical @stimulin@ gene $ith t$o exons encoding a protein of 1.. amino acids
is found to ha&e abnormal expression in cell culture. /hich of the follo$ing mutations
$ould produce a :..%bp stimulin m2NA and a 155Jamino acid peptide that reacts $ith
antibodies to stimulin protein0
A. )plice junction mutation pre&enting 2NA splicing
!. =rame%shift mutation in codon K+
". )ilent point mutation in the third nucleotide of codon K:.
D. Nonsense mutation at codon K+
#. Deletion of exon 1
The answer is: A
)plice junction mutations $ill theoretically produce a larger m2NA unless the m2NA is
unstable8 the larger protein may ha&e abnormal function but retain peptide regions that
react $ith antibody to the authentic protein. Nucleotide insertions or deletions other than
multiples of 5 alter the reading frame of the code and scramble the amino acid sequence
distal to the frame%shift mutation. )uch altered m2NAs may be of increased or smaller
si6e depending on their stability as may the translated protein depending on the
presence of stop codons $ithin the shifted reading frame. 7nly the protein upstream of
the frame%shift mutation retains immune cross%reacti&ity and normal function. 3oint
mutations (nucleotide substitutions) may ha&e substantial functional impact if the altered
codon results in an amino acid substitution. ;f no amino acid substitution occurs they are
called silent mutations.
11.
Template%directed 2NA synthesis occurs in $hich of the follo$ing0
A. 3oint mutation
!. Triplet repeat expansion
". ;nitiation of the polymerase chain reaction
D. #xpression of oncogenes
#. 2epair of thymine dimmers
The ans$er isG D
7ncogenes are cancer%producing genes. They are closely related to normal cellular genes
and are often tyrosine *inases gro$th factors or receptors for gro$th factors. The
expression of oncogenes leads to the translation and e&entual transcription of the protein
product of the oncogene. Thus template%directed 2NA synthesis but not DNA synthesis
occurs during the expression of oncogenes. ;n contrast template%directed DNA synthesis
rather than 2NA synthesis occurs during the repair of thymine dimers the polymerase
chain reaction the functioning of the replication for* and the gro$th of 2NA tumor
&iruses. ;n the final stages of the repair of thymine dimers once the dimer has been
excised DNA polymerase ; enters the gap to carry out template%directed synthesis. ;n
functioning of the replication for* DNA polymerase ;;; holoen6yme carries out synthesis
of DNA during replication. Template%directed DNA synthesis is required for the gro$th
of 2NA tumor &iruses (retro&iruses). 7nce released into the host cytoplasm retro&iral
2NA synthesi6es both the positi&e and minus strands of DNA using re&erse
transcriptase. This unique en6yme cataly6es the initial 2NA%directed DNA synthesis
hydrolysis of 2NA and then DNA%directed DNA synthesis. The ne$ly formed &iral
DNA duplex integrates into the host cell DNA prior to transcription. ;n this form the
retro&irus is inherited by daughter host cells. The polymerase chain reaction is a method
of amplifying the amount of DNA in a sample or of enriching particular DNA sequences
in a population of DNA molecules. ;n the polymerase chain reaction oligonucleotides
complementary to the ends of the desired DNA sequence are used as primer for multiple
rounds of template%directed DNA synthesis.
1+.
/hich of the follo$ing most correctly describes mammalian messenger 2NAs0
A. They are usually transcribed from both DNA strands
!. They are normally double%stranded
". Their content of uridine equals their content of adenine
D. They ha&e an o&erall negati&e charge at neutral p1
#. Their ratio of ribose to purine bases equals 1
The ans$er isG D
At a physiologic p1 of B., m2NAs (li*e DNA) are polyanionic o$ing to the negati&ely
charged phosphate hydroxyl groups. (ammalian m2NAs are synthesi6ed from DNA as
single%stranded linear molecules. !ecause they are not double%stranded the
concentrations of the different bases in m2NA are &ariable rather than exhibiting the A F
T and ' F " pattern of double%stranded DNA (A does not equal C in m2NA). The
hybridi6ation of 2NA $ith its complementary template DNA is antiparallel. ;n both DNA
and 2NA sugar units equal base units equal phosphate units. 1o$e&er their bases
consist of pyrimidines as $ell as purines.
15.
The $estern blot uses $hat type of probe0
A. Antibody
!. m2NA
". 3roducts of polymerase chain reaction (3"2)
D. t2NA
#. (utant and normal oligonucleotides
=. r2NA
'. cDNA clone
The ans$er isG A
;n an expression library cDNA clones are screened on the basis of their ability to direct
bacterial synthesis of a foreign protein of interest. 2adioacti&e antibodies specific to this
protein can be used to identify the colonies of bacteria that contain the cDNA &ector. As
$as the case for probing genomic libraries bacteria gro$n on a master plate are blotted
onto a nitrocellulose replica plate and then lysed. The released proteins may then be
labeled $ith 1+:; antibodies. ;n contrast northern blotting can be used to identify 2NA
molecules separated by gel electrophoresis. ;n northern blotting 2NA molecules
separated by gel electrophoresis can be identified by hybridi6ation $ith probe DNA
follo$ing transfer to nitrocellulose. (utant and $ild%type oligonucleotides can be used as
probes to analy6e polymerase chain reaction products. "on&ersely the products of
polymerase chain reaction can be used to analy6e cDNA libraries.
1,.
1o$ many high%energy phosphate%bond equi&alents are utili6ed in the process of
acti&ation of amino acids for protein synthesis0
A. Lero
!. 7ne
". T$o
D. Three
#. =our
The ans$er isG "
AT3 is required for the esterification of amino acids to their corresponding t2NAs. This
reaction is cataly6ed by the class of en6ymes *no$n as aminoacyl%t2NA synthetases.
#ach one of these en6ymes is specific for one t2NA and its corresponding amino acid.
amino acid I t2NA I AT3 aminoacyl%t2NA I A(3 I 33i
As $ith most AT3 hydrolysis reactions that release pyrophosphate pyrophosphatase
quic*ly hydroly6es the product to 3i $hich ma*es the reaction essentially irre&ersible.
)ince AT3 is hydroly6ed to A(3 and 33i during the reaction by con&ention the
equi&alent of t$o high%energy phosphate bonds is utili6ed.
1:.
An immigrant family from rural (exico brings their 5%month%old child to the emergency
room because of $histling inspiration (stridor) and high fe&er. The childAs physician is
perplexed because the throat examination sho$s a gray membrane almost occluding the
larynx. A senior physician recogni6es diphtheria no$ rare in immuni6ed populations.
The child is intubated antitoxin is administered and antibiotic therapy is initiated.
Diphtheria toxin is often lethal in unimmuni6ed persons because it
A. ;nhibits initiation of protein synthesis by pre&enting the binding of 'T3 to the
,.) ribosomal subunit
!. !inds to the signal recognition particle receptor on the cytoplasmic face of the
endoplasmic reticulum receptor
". )huts off signal peptidase
D. !loc*s elongation of proteins by inacti&ating elongation factor + (#=%+ or
translocase)
#. "auses deletions of amino acid by speeding up the mo&ement of peptidyl%t2NA
from the A site to the 3 site
The ans$er isG D
The gene that produces the deadly toxin of "orynebacterium diphtheriae comes from a
lysogenic phage that gro$s in the bacteria. 3rior to immuni6ation diphtheria $as the
primary cause of death in children. The protein toxin produced by this bacterium inhibits
protein synthesis by inacti&ating elongation factor + (#=%+ or translocase). Diphtheria
toxin is a single protein composed of t$o portions (A and !). The ! portion enables the A
portion to translocate across a cell membrane into the cytoplasm. The A portion cataly6es
the transfer of the adenosine diphosphate ribose unit of NAD1 to a nitrogen atom of the
diphthamide ring of #=%+ thereby bloc*ing translocation. Diphthamide is an unusual
amino acid residue of #=%+.
19.
(ethionylJtransfer (t) 2NA is used for initiation of protein synthesis by $hich of the
follo$ing0
A. "hloroplast ribosomes
!. #u*aryotic mitochondrial ribosomes
". #u*aryotic cytoplasmic ribosomes
D. !acterial ribosomes
#. !acterial cytoplasm
The ans$er isG "
(ethionyl%t2NA is the special t2NA used in eu*aryotes for initiation. ;nitiation of
protein synthesis by bacterial mitochondrial and chloroplast ribosomes requires N%
formylmethionyl%t2NA. The mitochondria of eu*aryotic cells are similar to bacteria in
the si6e of their ribosomal 2NAs (+5) and 19)) and their mechanisms for protein
synthesis. (itochondrial and pro*aryotic ribosomes (including those of chloroplasts and
bacteria) use formylmethionyl%t2NA for initiation of protein synthesis and are sensiti&e
to inhibitors li*e streptomycin tetracycline and chloramphenicol that ha&e little effect on
eu*aryotic cells. The latter drugs are useful as antibiotics in animals and humans since
they inhibit bacteria but do not gain entry into mitochondria.
1B.
/hich of the follo$ing is required for certain types of eu*aryotic protein synthesis but
not for pro*aryotic protein synthesis0
A. 2ibosomal 2NA
!. (essenger 2NA
". )ignal recognition particle
D. 3eptidyl transferase
#. 'T3
The answer is: C
)ignal recognition particles ()23s) recogni6e the signal sequence on the N%terminal end
of proteins destined for the lumen of the endoplasmic reticulum (#2). )23 binding
arrests translation and an )23 receptor facilitates import of the )23 ribosome and
nascent protein into the #2 lumen. A signal peptidase remo&es the signal sequence from
the protein $hich may remain in the membrane or be routed for secretion. "ommon to
both eu*aryotic and pro*aryotic protein synthesis is the requirement for AT3 to acti&ate
amino acids. The acti&ated aminoacyl%t2NAs then interact $ith ribosomes carrying
m2NA. 3eptidyl transferase cataly6es the formation of peptide bonds bet$een the free
amino group of acti&ated aminoacyl%t2NA on the A site of the ribosome and the esterified
carboxyl group of the peptidyl%r2NA on the 3 site8 the liberated r2NA remains on the 3
site.
1E.
A major obstacle to gene therapy in&ol&es the difficulty of homologous gene
replacement. /hich of the follo$ing strategies addresses this issue0
A. A recombinant &ector contains complementary DNA sequences that $ill facilitate
site%specific recombination
!. A recombinant &ector expresses antisense nucleotides that $ill hybridi6e $ith the
targeted m2NA
". A recombinant &ector replaces inessential &iral genes $ith a functional human
gene
D. A recombinant &ector transfects patient cells $hich are returned to the patient
#. A recombinant &ector contains DNA sequences that target its expressed protein to
lysosomes
The answer is: A
"hallenges for gene therapy include the construction of recombinant &iral genomes that
can propagate the replacement gene (gene constructs or &ectors) deli&ery of the altered
gene to the appropriate tissues (gene targeting) and recombination at the appropriate
locus so that replacement of the defecti&e gene is achie&ed (site%specific recombination).
The latter step positions DNA sequences in the &ector so that the replacement gene pairs
and recombines precisely $ith homologous DNA in the nati&e gene. #x &i&o transfection
(introduction of &ector DNA into patient cells outside the body) is an ideal method for
gene targeting if the engineered cells can repopulate the tissueMorgan in question.
Transfection of bone marro$ stem cells $ith a functional adenosine deaminase gene
follo$ed by bone marro$ transplantation bac* to the patient has been successful in
restoring immunity to children $ith se&ere deficiency. #&en $hen tissue targeting and
precise gene replacement are feasible mimic*ing the appropriate patterns of gene
expression can be a substantial barrier to gene therapy. ;njection of deficient en6ymes
into serum (en6yme therapy) has been successful in disorders such as 'aucherAs disease
<+51... (storage of lipids in the spleen and bone)? and ta*es ad&antage of cellular
path$ays that target en6ymes to lysosomes.
1-.
A family in $hich se&eral indi&iduals ha&e arthritis and detached retina is diagnosed $ith
)tic*ler syndrome. The locus for )tic*ler syndrome has been mapped near that for type ;;
collagen on chromosome 1+ and mutations in the "7>+A1 gene ha&e been described in
)tic*ler syndrome. The family became interested in molecular diagnosis to distinguish
normal from mildly affected indi&iduals. /hich of the follo$ing results $ould be
expected in an indi&idual $ith a promoter mutation at one "7>+A1 gene locus0
A. /estern blotting detects no type ;; collagen chains
!. )outhern blotting using intronic restriction sites yields normal restriction fragment
si6es
". 2e&erse transcriptaseJpolymerase chain reaction (2T%3"2) detects one%half
normal amounts of "7>+A1 m2NA in affected indi&iduals
D. =luorescent in situ hybridi6ation (=;)1) analysis using a "7>+A1 probe detects
signals on only one chromosome 1+
#. DNA sequencing re&eals a single nucleotide difference bet$een homologous
"7>+A1 exons
The ans$er isG "
After the locus responsible for a genetic disease is mapped to a particular chromosome
region @candidate@ genes can be examined for molecular abnormalities in affected
indi&iduals. The connecti&e tissue abnormalities in )tic*ler syndrome (1.E5..) ma*e the
"7>+A1 collagen locus an attracti&e candidate for disease mutations prompting analysis
of "7>+A1 gene structure and expression. /estern blotting detects gene alterations that
interfere $ith protein expression $hile use of the re&erse transcriptaseJpolymerase chain
reaction (2T%3"2) detects alterations in m2NA le&els. #ach analysis should detect one%
half the respecti&e amounts of "7>+A1 protein or m2NA in the case of a promoter
mutation that abolishes transcription of one "7>+A1 allele. )outhern blotting detects
nucleotide changes that alter DNA restriction sites but this is relati&ely insensiti&e unless
large portions of the gene are deleted. =luorescent in situ hybridi6ation (=;)1) analysis
using DNA probes from the "7>+A1 locus is a sensiti&e method for detecting deletions
of the entire locus and DNA sequencing of the entire gene pro&ides the gold standard for
detecting any alteration in the regulatory or coding sequences. Nucleotide sequence
changes are still subject to interpretation since they may represent polymorphisms that
do not alter gene function. 3opulation studies andMor in &itro studies of gene expression
are often needed to discriminate DNA polymorphisms from mutations that disrupt gene
function. =or any autosomal locus the interpretation of molecular analyses is
complicated by the presence of t$o homologous copies of the gene.
+..
'yrate atrophy (+:EEB.) is a rare autosomal recessi&e genetic disorder caused by a
deficiency of ornithine aminotransferase. Affected indi&iduals experience progressi&e
chorioretinal degeneration. The gene for ornithine aminotransferase has been cloned its
structure has been determined and mutations in affected indi&iduals ha&e been
extensi&ely studied. /hich of the mutations listed belo$ best fits $ith test results
sho$ing normal )outhern blots $ith probes from all ornithine aminotransferase exons but
absent en6ymatic acti&ity0
A. Duplication of entire gene
!. T$o%*b deletion in coding region of gene
". T$o%*b insertion in coding region of gene
D. Deletion of entire gene
#. (issense mutation
The answer is: E
(issense mutations $hich cause the substitution of one amino acid for another may
significantly alter the function of the resultant protein $ithout altering the si6e of DNA
restriction fragments detected by )outhern blotting. ;n this case northern blot results
$ould most li*ely also be normal. )ingle%base changes may also result in nonsense
mutations. >arge insertions or deletions in the exon or coding regions of the gene alter the
)outhern blot pattern and usually ablate the acti&ity of one gene copy. ;n the case of an
autosomal locus li*e that for ornithine aminotransferase the homologous allele remains
acti&e and gi&es :.N en6yme acti&ity (hetero6ygote or carrier range $ith a normal
phenotype). )imilar effects on en6yme acti&ity $ould be predicted from complete gene
deletions at one locus $hile a duplication might produce 1:.N or :.N of normal
en6yme acti&ity depending on the status of promoter sites.
+1.
1urlerAs syndrome (+:+E..) is caused by a deficiency of >%iduronidase an en6yme
normally expressed in most human cell types. ;t $as demonstrated by Neufeld that
exogenous >%iduronidase could be ta*en up by deficient cells &ia a targeting signal that
directed the en6yme to its normal lysosomal location. /hich of the follo$ing therapeutic
strategies $ould be the most realistic and efficient mode of therapy0
A. 'erm%line gene therapy
!. 1eterologous bone marro$ transplant
". ;nfection $ith a disabled adeno&irus &ector that carries the >%iduronidase gene
D. ;njection $ith >%iduronidase purified from human li&er
#. Autologous bone marro$ transplant after transfection $ith a &irus carrying the >%
iduronidase gene
The answer is: B
All of the modes of therapy are theoretically possible and en6yme therapy (i.e. injection
of purified en6yme) has been successful in se&eral lysosomal deficiencies particularly
those in $hich the central ner&ous system is not affected <i.e. 'aucherAs disease
(+51...)?. Cnfortunately antibodies frequently de&elop to the injected en6yme and limit
the term of successful en6yme deli&ery. 1eterologous bone marro$ transplant preferably
from a related donor offers the most realistic and effecti&e therapy since the graft
pro&ides a permanent source of en6yme. !one marro$ transplants do ha&e a 1.N
mortality ho$e&er and the en6yme diffuses poorly into the central ner&ous system.
)omatic gene therapy (i.e. deli&ery of en6yme to somatic cells &ia &iral &ectors or
transfected tissue) is no$ possible8 ho$e&er targeting of the gene product to appropriate
tissues and organelles is still a problem. Transfected autologous bone marro$ transplant
(i.e. marro$ from the patient) has been used in a fe$ cases of adenosine deaminase
deficiency an immune disorder affecting lymphocytes. 'erm%line gene therapy requires
the insertion of functional genes into gametes or blastomeres of early embryos prior to
birth. The potential for embryonic damage lac* of *no$ledge regarding de&elopmental
gene control and ethical contro&ersies regarding selecti&e breeding or embryo
experimentation ma*e germ%line therapy unrealistic at present.
++.
/hich of the follo$ing mutations is most li*ely to be lethal0
A. )ubstitution of adenine for cytosine
!. )ubstitution of cytosine for guanine
". )ubstitution of methylcytosine for cytosine
D. Deletion of three nucleotides
#. ;nsertion of one nucleotide
The answer is: E
;nsertion of one extra nucleotide causes a frame%shift mutation and mistranslation of all
the m2NA transcribed from beyond that point in the DNA. All the other mutations cited
in the question usually cause an error in the identity of only one amino acid (choice a or
b) remo&al of one amino acid from the sequence (choice d) or no error at all in the
amino acid sequence (choice c). There is a chance that the mutations in choices a or b
$ill gi&e a @nonsense@ or chain%terminator mutation and this is about as li*ely to be
lethal as is a frame shift.
+5.
;n the follo$ing partial sequence of m2NA a mutation of the template DNA results in a
change in codon -1 to CAA. /hat type of mutation is it0
EE E- -. -1 -+ -5 -,
'C" 'A" "A' CA' ''" CAA ""'
A. (issense
!. )ilent
". Nonsense
D. )uppressor
#. =rame shift
The answer is: B
The replacement of the codon CA' $ith CAA $ould be a silent mutation since both
codons are @stop@ signals. Thus transcription $ould cease $hen either triplet $as
reached. There are three termination codons in m2NAG CA' CAA and C'A. These are
the only codons that do not specify an amino acid. A missense or a substitution mutation
is the con&erting of a codon specifying one amino acid to another codon specifying a
different amino acid. A nonsense mutation con&erts an amino acid codon to a termination
codon. A suppression counteracts the effects of another mutation at another codon. The
addition or deletion of nucleotides results in a frame%shift mutation.
+,.
/hich one of the follo$ing causes a frame%shift mutation0
A. Transition
!. Trans&ersion
". Deletion
D. )ubstitution of purine for pyrimidine
#. )ubstitution of pyrimidine for purine
The ans$er isG "
3oint mutations that are frame%shift mutations put the normal reading frame out of
register by one base pair. The insertion of an extra base pair or the deletion of one or
more base pairs falls into this category. Transitions and trans&ersions are not frame%shift
mutations8 they are substitutions of one base pair for another. )ubstitutions are the most
common type of mutation. ;n transitions a purine is replaced by a purine or a pyrimidine
by a pyrimidine. ;n trans&ersions a purine is replaced by a pyrimidine or &ice &ersa. ;t
has been suggested that transitions occur spontaneously o$ing to the tautomeric changes
in base%hydrogen%bond locations. Trans&ersions can be caused by defecti&e DNA
polymerases.
+:.
/hich of the follo$ing regulators are said to act in @cis@0
A. The lac repressor and mammalian transcription factors
!. The lac repressor and the lac operator
". The lac operator and mammalian enhancers
D. The lac operator and mammalian transcription factors
#. (ammalian transcription factors and enhancers
The ans$er isG "
"ertain regulatory elements act on genes on the same chromosome (@cis@) $hile others
can regulate genes on the opposite chromosome (@trans@). The terminology ma*es
analogy to carbon%carbon double bonds $here t$o modifying groups may both be abo&e
or belo$ the bond (cis) or opposite it (trans). "is regulatory elements li*e the lac operator
and promoter or mammalian enhancers are usually DNA sequences (regulatory
sequences) adjoining or $ithin the regulated gene. Trans regulatory elements li*e the lac
repressor protein or mammalian transcription factors are usually diffusible proteins
(regulatory factors) that can interact $ith adjoining target genes or $ith target genes on
other chromosomes. "lassification of bacterial elements as cis or trans requires mating
experiments $here portions of a second chromosome are introduced by transfection ($ith
bacteriophage) or conjugation ($ith other bacteria). The distinction bet$een cis and trans
is fundamental for understanding ho$ regulators $or*.
+9.
/hich of the structural domains of mammalian regulatory factors may be called
intracellular receptors0
A. 2esponse elements
!. Antirepressor domains
". Transcription%acti&ating domains
D. >igand%binding domains
#. DNA%binding domains
The answer is: D
(ammalian regulatory factors are much more di&erse than those of bacteria possessing
se&eral types of structural domains. Acti&ators of transcription such as steroid hormones
may enter the cell and bind to regulatory factors at specific sites called ligand%binding
domains8 these intracellular @receptors@ are analogous to ' proteinJlin*ed membrane
receptors that extend into the extracellular space. 2esponse elements are not regulatory
factors but DNA sequences near the transcription site for certain types of genes (e.g.
steroid%responsi&e and heat shoc*Jresponsi&e genes). 2egulatory factors interact $ith
specific DNA sequences through their DNA%binding domains and $ith other regulatory
factors through transcription%acti&ating domains. )ome regulatory factors ha&e
antirepressor domains that counteract the inhibitory effects of chromatin proteins
(histones and nonhistones).
+B.
The proopiomelanocortin (37(") gene encodes se&eral regulatory proteins that affect
pituitary function. ;n different brain regions proteins encoded by this gene ha&e different
carboxy%terminal peptides. /hich of the follo$ing best explains the regulatory
mechanism0
A. 37(" transcription is regulated by different factors in different brain regions
!. 37(" translation elongation is regulated by different factors in different brain
regions
". 37(" transcription has different enhancers in different brain regions
D. 37(" protein undergoes different protein processing in different brain regions
#. 37(" protein forms different allosteric complexes in different brain regions
The answer is: D
The 37(" gene pro&ides a mammalian example in $hich se&eral proteins are deri&ed
from the same 2NA transcript. Cnli*e the polycistronic m2NA of the bacterial lactose
operon mammalian cells generate se&eral m2NAs or proteins from the same gene by
&ariable protein processing or by alternati&e splicing. Dariable protein processing
preser&es the peptide products of some gene regions but degrades those from others.
Alternati&e splicing $ould often produce proteins composed of different exon
combinations $ith the same terminal exon and carboxy%terminal peptide but could
remo&e the terminal exon in some proteins and produce different "%terminal peptides.
Different transcription factors or enhancers in different brain regions could regulate the
total amounts of 37(" gene transcript but not the types of protein produced. #longation
of protein synthesis in&ol&es 'T3 clea&age but is not differentially regulated in
mammalian tissues.
+E.
T$o boys $ith mental disability are found to ha&e mutations in a gene on the O
chromosome that has no homology $ith globin genes. !oth are also noted to ha&e
deficiency of %globin synthesis $ith thalassemia. /hich of the follo$ing is the best
explanation for their phenotype0
A. The mutation disrupted an enhancer for an %globin pseudogene
!. The mutation disrupted an O%encoded transcription factor that regulates the
%globin loci
". There is a second mutation that disrupts an enhancer near the %globin gene
D. There is a DNA rearrangement that joins the mutated O chromosome gene $ith an
%globin gene
#. There is a second mutation that disrupts the promoter of an %globin gene
The ans$er isG !
The boys ha&e an O%lin*ed recessi&e condition called thalassemiaMmental retardation or
AT2%O syndrome (5.-:1.). The O%encoded gene has an un*no$n function in brain as
$ell as being a factor that regulates %globin gene transcription. ;n order to affect all four
%globin genes the O%encoded gene must produce a trans%acting factor8 second mutations
altering enhancers or promoters $ould be cis%acting and affect only one %globin gene.
3seudogenes are functionless gene copies so altered expression $ould not influence
%globin chain synthesis.
+-.
A middle%aged man presents $ith a mar*edly enlarged tonsil and recurrent infections
$ith serum immunoglobulin deficiency. "hromosome analysis demonstrates a
translocation bet$een the immunuglobulin hea&y chain locus on chromosome 1, and an
unidentified gene on chromosome E. /hich of the follo$ing is the most li*ely cause of
his phenotype0
A. The translocation has deleted constant chain exons on chromosome 1, and
pre&ented hea&y chain class s$itching
!. The translocation has deleted the inter&al containing di&ersity (D) and joining (P)
regions
". The translocation has acti&ated a tumor%promoting gene on chromosome E
D. The translocation has deleted the hea&y chain constant chain " so that &irgin !
cells cannot produce ;g( on their membranes
#. The translocation has deleted an immunoglobulin transcription factor gene on
chromosome E
The answer is: C
This case is an example of !ur*ittAs lymphoma $hich may affect the tonsils or other
lymphoid tissues. The translocation places the myc oncogene on chromosome E
do$nstream of the &ery acti&e hea&y chain locus on chromosome 1, acti&ating myc gene
expression in ! cells and their deri&ati&es. The translocation is li*ely an aberrant form of
the normal DNA rearrangements that generate unique hea&y chain genes in each ! cell.
The translocation joins one chromosome E to one chromosome 1, lea&ing their
homologues unaffected. The cause for the phenotype must therefore be trans-acting
since cis-acting effects $ould pertain only to the translocated loci and not affect the
homologous untranslocated loci. Acti&ation of a tumor%promoting gene (oncogene) on
chromosome E could produce an enlarged tonsil $hile underacti&ity of immunoglobulin
production due to one%half expression could decrease immune function but $ould not
completely ablate the processes in choices a b d and e. At the genetic le&el trans-acting
e&ents are autosomal dominant in that one of the t$o homologous loci is abnormal and
produces a phenotype. (utations of cis-acting e&ents must disrupt both homologous loci
to produce phenotypes ma*ing them autosomal recessi&e at the genetic le&el.
5..
A +%day%old neonate becomes lethargic and uninterested in breast%feeding. 3hysical
examination re&eals tachypnea (rapid breathing) $ith a normal heartbeat and breath
sounds. ;nitial blood chemistry &alues include normal glucose sodium potassium
chloride and bicarbonate (1"75
J
) le&els8 initial blood gas &alues re&eal a p1 of B.:5
partial pressure of oxygen (37+) normal at 1.5 mm1g and partial pressure of carbon
dioxide (3"7+) decreased at +B mm1g. /hich of the follo$ing is the most appropriate
treatment0
A. Administer al*ali to treat metabolic acidosis
!. Administer al*ali to treat respiratory acidosis
". Decrease the respiratory rate to treat metabolic acidosis
D. Decrease the respiratory rate to treat respiratory al*alosis
#. Administer acid to treat metabolic al*alosis
The answer is: D
Tachypnea in term infants may result from brain injuries or metabolic diseases that
irritate the respiratory center. The increased respiratory rate remo&es (@blo$s off@) carbon
dioxide from the lung al&eoli and lo$ers blood "7+ forcing a shift in the indicated
equilibrium to$ard the leftG

"arbonic acid (1+"7+) can be ignored because negligible amounts are present at
physiologic p1 lea&ing the equilibriumG
The left$ard shift to replenish exhaled "7+ decreases the hydrogen ion (1
I
)
concentration and increases the p1 (
%
log1.<1
I
?) to produce al*alosis (blood p1 abo&e the
physiologic norm of B.,). This respiratory al*alosis is best treated by diminishing the
respiratory rate to ele&ate the blood <"7+? force the abo&e equilibrium to the right
ele&ate the <1
I
? and decrease the p1. The ne$born does not ha&e acidosis defined as a
blood p1 belo$ B., either from excess blood acids (metabolic acidosis) or from
increased <"7+? (respiratory acidosis). The baby also does not ha&e metabolic al*alosis
caused by loss of hydrogen ion from the *idney (e.g. $ith defecti&e tubular filtration) or
stomach (e.g. $ith se&ere &omiting).
51.
A ne$born $ith tachypnea and cyanosis (bluish color) is found to ha&e a blood p1 of B.1.
A serum bicarbonate is measured as 1+ mM, but the blood gas machine that $ould
determine the partial pressures of oxygen (37+) and carbon dioxide (3"7+) is bro*en.
2ecall the pKa of 9.1 for carbonic acid (reflecting the 1"75
J
M"7+ equilibrium in blood)
and the fact that the blood "7+ concentration is equal to the 3"7+ in mm1g (normal
&alue F ,. mm1g) multiplied by ...5. /hich of the follo$ing is the most appropriate
treatment0
A. Administer oxygen to impro&e tissue perfusion and decrease metabolic acidosis
!. Administer oxygen to decrease respiratory acidosis
". ;ncrease the respiratory rate to treat respiratory acidosis
D. Decrease the respiratory rate to treat respiratory acidosis
#. Administer medicines to decrease renal hydrogen ion excretion
The answer is: A
The equilibrium bet$een an acid and its conjugate base is defined by the 1enderson%
1asselbalch equationG
in the case of carbonic acid. Note that "7+ is the effecti&e acid and 1"75
J
the conjugate
base for carbonic acid due to its complete dissociation in $ater.
'i&en a p1 of B.1 in the cyanotic ne$born then B.1 J 9.1 F 1 F log (1.) F log <1"75
J
?M
<"7+? F log <1"75
J
?M...5 H 3"7+. )ince the <1"75
J
? is 1+ mM, the 3"7+ H ...5 must
be 1.+ mM and the 3"7+ ,. mm1g. This normal calculated &alue for 3"7+ means that the
baby must ha&e metabolic acidosis a common accompaniment of hypoxia (lo$ 37+) that
can be treated by pro&iding oxygen or administering al*ali to ameliorate the acidosis. ;f
the baby had respiratory acidosis the 3"7+ $ould be ele&ated8 this $ould be treated by
increasing the respiratory rate to blo$ off "7+. 2enal treatment of acidosis $ould require
increasing acid excretion or al*ali retention. The lungs compensate acidosis $ith
increased breathing rates or tidal &olumes to blo$ off "7+ and increase p1 the *idneys
by retaining 1"75
J
. The lungs can compensate al*alosis some$hat by decreasing
breathing rates or &olumes to retain "7+ (and decrease oxygenation $ithin limits) the
*idneys by increasing excretion of 1"75
J
.
5+.
A diabetic teenager is found to ha&e a p1 of B.1 and normal electrolyte le&els (Na
I
F 1,.
mM, Q
I
F , mM, "l
J
F 1.5 mM) except for a bicarbonate of 11 mM (normal ++ to +E
mM). The urine tests positi&e for *etone bodies mostly due to acetoacetic acid and
acetoacetate ("15"F7"1+"771 and "15"F7"1+"77
J
) $hich ha&e a pK of ,.E. ;n
this case it is assumed that acetoacetate is the only significant anion in the blood besides
chloride and that each acetoacetate anion binds and remo&es one sodium cation during
excretion by the *idney. 'i&en that the patient has a normal glomerular filtration rate of
about B > of blood per hour $ithout any retention of acetoacetateMacetoacetic acid the
rates of sodium acetoacetate and acetoacetic acid loss $ill be
A. 1. mmolMh of each species
!. :. mmolMh of sodium and acetoacetate &irtually no acetoacetic acid excretion
". 1.. mmolMh of sodium and acetoacetic acid &irtually no acetoacetate excretion
D. +.. mmolMh of sodium and acetoacetate &irtually no acetoacetic acid excretion
#. 5.. mmolMh of each species
The answer is: D
The sum of the major cations in blood (Na
I
plus Q
I
) minus the sum of the major anions
(1"75
J
plus "l
J
) is called the anion gap (filled by phosphate ions negati&ely charged
proteins etc.). An anion gap o&er +. suggests the presence of an abnormal anion such as
acetoacetate $hich occurs in diabetics. The anion gap in this teenager is ele&ated at 5.
(1,. I ,) J (1.5 I 11). Assuming that all of the gap is made up by acetoacetate anion
then B > H 5. mmolM> F +1. mmol of acetoacetate and +1. mmol of sodium excreted per
hour. #&en $ith acidosis and a p1 of B.1 &irtually all of the acetoacetic acid (pK ,.E) is
present as acetoacetate and less than 1N is excreted as acetoacetic acid. To calculate the
exact amount of acetoacetic acid present the 1enderson%1asselbalch equation rearranges
to (B.1 J ,.E) F +.5 F log<base?M<acid?8 <"15"F7"1+"77
J
?M<"15"F7"1+"771? F
antilog +.5 F 1.+. >ess than ..5 mmol of acetoacetic acid is excreted for each liter of
blood filtered through the *idney.
55.
/hich of the combinations of laboratory results belo$ indicates compensated metabolic
al*alosis0
A. >o$ 3"7+ normal bicarbonate high p1
!. >o$ 3"7+ lo$ bicarbonate lo$ p1
". Normal 3"7+ lo$ bicarbonate lo$ p1
D. 1igh 3"7+ normal bicarbonate lo$ p1
#. 1igh 3"7+ high bicarbonate high p1
The answer is: E
3ure metabolic acidosis (choice c) or pure metabolic al*alosis exhibits abnormal
bicarbonate and normal lung function. 3ure respiratory acidosis (choice d) or al*alosis
(choice a) is associated $ith normal renal function (and normal blood acids) $ith a
normal bicarbonate and abnormal 3"7+. Thus choices b and e must in&ol&e
compensation since both the 3"7+ and bicarbonate are abnormal. "hoice e must
represent compensated metabolic al*alosis since the 3"7+ is high4if it $ere
compensated respiratory acidosis $ith a high 3"7+ the p1 $ould be lo$.
5,.
The p1 of body fluids is stabili6ed by buffer systems. /hich of the follo$ing compounds
is the most effecti&e buffer at physiologic p10
A. Na+137, pQa: 1+.5+
!. N1,71 pQa: -.+,
". Na1+37, pQa: B.+1
D. "15"7+1 pQa: ,.B,
#. "itric acid pQa: 5..-
The ans$er isG "
;n any fluid maximum buffering action is achie&ed by the acid $hose pQa most nearly
approximates the p1 of the fluid. 3hysiologic p1 is about B., so that among those
buffers listed in the question Na1+37, is the most effecti&e.
5:.
A child presents $ith se&ere &omiting dehydration and fe&er. ;nitial blood studies sho$
acidosis $ith a lo$ bicarbonate and an anion gap (the sum of sodium plus potassium
minus chloride plus bicarbonate is ,. and larger than the normal +. to +:). 3reliminary
results from the blood amino acid screen sho$ t$o ele&ated amino acids both $ith
nonpolar side chains. A titration cur&e performed on one of the ele&ated species sho$s
t$o ioni6able groups $ith approximate pKs of + and -.:. The most li*ely pair of ele&ated
amino acids consists of
A. Aspartic acid and glutamine
!. 'lutamic acid and threonine
". 1istidine and &aline
D. >eucine and isoleucine
#. 'lutamine and isoleucine
The answer is: D
>eucine and isoleucine ha&e nonpolar methyl groups as side chains. As for any amino
acid titration cur&es obtained by noting the change in p1 o&er the range of 1 to 1, $ould
sho$ a pK of about + for the primary carboxyl group and about -.: for the primary amino
group8 there $ould be no additional pK for an ioni6able side chain. 2ecall that the pK is
the point of maximal buffering capacity $hen the amounts of charged and uncharged
species are equal (see ans$er to question 1.,). Aspartic and glutamic acids (second
carboxyl group) histidine (imino group) and glutamine (second amino group) all ha&e
ioni6able side chains that $ould gi&e an additional pK on the titration cur&e. The li*ely
diagnosis here is maple syrup urine disease $hich in&ol&es ele&ated isoleucine leucine
and &aline together $ith their *etoacid deri&ati&es. The *etoacid deri&ati&es cause the
acidosis and the fe&er suggests that the metabolic imbalance $as $orsened by an
infection.
59.
/hich of the hemoglobin designations belo$ best describes the relationship of subunits
in the quaternary structure of adult hemoglobin0
A. ( 1% +)( 1% +) B. 1% +% 5% , C. % % % D. ( 1% +% 5% 1) E. ( 1%
1)%( +% +)
The answer is: E
Adult hemoglobin or hemoglobin A is composed of four polypeptide chains. T$o of the
chains are chains and t$o are chains. The chains are held together by nonco&alent
interactions. The hemoglobin tetramer can best be represented as being composed of t$o
dimers each containing the t$o different polypeptides. Thus the designation ( 1% 1) ( +%
+) $hich refer to dimers 1 and + respecti&ely is the most correct $ay to refer to the
quaternary structure of adult hemoglobin. 1ydrophobic interactions are thought to be the
main nonco&alent interactions holding all four polypeptides together.
5B.
!lood is dra$n from a child $ith se&ere anemia and the hemoglobin protein is degraded
for peptide and amino acid analysis. 7f the follo$ing results $hich change in
hemoglobin primary structure is most li*ely to correlate $ith the clinical phenotype of
anemia0
A. ile%leu%&al to ile%ile%&al
!. leu%glu%ile to leu%&al%ile
". gly%ile%gly to gly%&al%gly
D. gly%asp%gly to gly%glu%gly
#. &al%&al%&al to &al%leu%&al
The answer is: B
3rimary protein structures denote the sequence of amino acids held together by peptide
bonds (carboxyl groups joined to amino groups to form amide bonds). The types of
amino acids then determine the secondary structure of peptide regions $ithin the protein
sometimes forming spiral helices or flat pleated sheets. These regional peptide
secondary structures then determine the o&erall three%dimensional tertiary structure of a
protein $hich is &ital for its function. Amino acid substitutions that alter the charge of an
amino acid side chain li*e the change from glutamic acid (charged carboxyl group) to
&aline (nonpolar methyl groups) in choice b are most li*ely to change the secondary and
tertiary protein structure. A change in hemoglobin structure can cause instability
decreased mean cellular hemoglobin concentration (("1") and anemia. A change from
glutamic acid to &aline at position 9 in the %hemoglobin chain is the mutation
responsible for sic*le cell anemia.
5E.
3arents bring in their +%$ee*%old child fearful that he has ingested a poison. They had
delayed disposing one of the childAs diapers and noted a blac* discoloration $here the
urine had collected. >ater they reali6ed that all of the childAs diapers $ould turn blac* if
stored as $aste for a day or so. Qno$ing that phenol groups can complex to form colors
$hich of the follo$ing amino acid path$ays are implicated in this phenomenon0
A. The phenylalanine tyrosine and homogentisate path$ay
!. The histidine path$ay
". The leucine isoleucine and &aline path$ay
D. The methionine and homocystine path$ay
#. The arginine and citrulline path$ay (urea cycle)
The answer is: A
>ac* of the en6yme homogentisate oxidase causes the accumulation of homogentisic
acid a metabolite in the path$ay of degradation of phenylalanine and tyrosine.
1omogentisate li*e tyrosine contains a phenol group. ;t is excreted in the urine $here it
oxidi6es and is polymeri6ed to a dar* substance upon standing. Cnder normal conditions
phenylalanine is degraded to tyrosine $hich is bro*en do$n through a series of steps to
fumarate and acetoacetate. The dar* pigment melanin is another end product of this
path$ay. Deficiency of homogentisate oxidase is called al*aptonuria (blac* urine) a mild
disease disco&ered by )ir Archibald 'arrod the pioneer of biochemical genetics.
'arrodAs geneticist colleague /illiam !ateson recogni6ed that al*aptonuria li*e nearly
all en6yme deficiencies exhibits autosomal recessi&e inheritance.
5-.
"ertain amino acids are not part of the primary structure of proteins but are modified
after translation. ;n scur&y $hich amino acid that is normally part of collagen is not
synthesi6ed0
A. 1ydroxytryptophan
!. 1ydroxytyrosine
". 1ydroxyhistidine
D. 1ydroxyalanine
#. 1ydroxyproline
The ans$er isG #
1ydroxyproline and hydroxylysine are not present in ne$ly synthesi6ed collagen. 3roline
and lysine residues are modified by hydroxylation in a reaction requiring the reducing
agent ascorbic acid (&itamin "). The en6ymes cataly6ing the reactions are prolyl
hydroxylase and lysyl hydroxylase. ;n scur&y $hich results from a deficiency of &itamin
" insufficient hydroxylation of collagen causes abnormal collagen fibrils. The $ea*ened
collagen in teeth bone and blood &essels causes tooth loss brittle bones $ith fractures
and bleeding tendencies $ith bruising and bleeding gums.
,..
A ne$born female has a large and distorted cranium short and deformed limbs and &ery
blue scleras ($hites of the eyes). 2adiographs demonstrate multiple limb fractures and
suggest a diagnosis of osteogenesis imperfecta (brittle bone disease). Analysis of type ;
collagen protein a triple helix formed from t$o 1 and one + collagen chains sho$s a
:.N reduction in the amount of type ; collagen in the babyAs s*in. DNA analysis
demonstrates the presence of t$o normal 1 alleles and one normal + allele. /hich of
the follo$ing is the best explanation of these results0
A. Deficiency of 1 collagen peptide synthesis
B. ;nability of 1 chains to incorporate into triple helix
C. Defecti&e 1 chains that interrupt triple helix formation
D. ;ncorporation of defecti&e + chains that cause instability and degradation of the triple
helix
E. A missense mutation that alters the synthesis of 1 chains
The answer is: D
"ollagen peptides assemble into helical tertiary structures that form quaternary triple
helices. The triple helices in turn assemble end to end to form collagen fibrils that are
essential for connecti&e tissue strength. 7&er 1: types of collagen contribute to the
connecti&e tissue of &arious organs including the contribution of type ; collagen to eyes
bones and s*in. The fact that only one of t$o + alleles is normal in this case implies that
a mutant + allele could be responsible for the disease (e&en if the + locus is on the O
chromosome since the baby is female $ith t$o O chromosomes). The mutant +
collagen peptide $ould be incorporated into half of the type ; collagen triple helices
causing a :.N reduction in normal type ; collagen. (A mutant 1 collagen peptide $ould
distort B:N of the molecules since t$o 1 peptides go into each triple helix.) The ability
of one abnormal collagen peptide allele to alter triple%helix structure $ith subsequent
degradation is $ell documented and colorfully named protein suicide or more properly a
dominant%negati&e mutation.
,1.
A child $ith tall stature loose joints and detached retinas is found to ha&e a mutation in
type ;; collagen. 2ecall that collagen consists of a repeating tripeptide motif $here the
first amino acid of each tripeptide is the same. /hich of the follo$ing amino acids is the
recurring amino acid most li*ely to be altered in mutations that distort collagen
molecules0
A. 'lycine B. 1ydroxyproline C. 1ydroxylysine D. Tyrosine E.
Tryptophan
The answer is: A
The primary structure of collagen peptides consists of repeating tripeptides $ith a gly-X-
Y motif $here gly is glycine and X and Y are any amino acid. The small "1+ group
connecting the amino and carboxyl groups of glycine contrasts $ith the larger connecting
groups and side chains of other amino acids. The small &olume of glycine molecules is
crucial for the helix secondary structure of collagen peptides. This in turn is necessary
for their tertiary helical structure and their assembly into quaternary tripeptide triple%
helix structures. The most se&ere clinical phenotypes caused by amino acid substitutions
in collagen peptides are those affecting glycine that pre&ent helix formation. The child
has a disorder called )tic*ler syndrome (1.E5..) that exhibits autosomal dominant
inheritance.
,+.
/hich of the follo$ing techniques for purification of proteins can be made specific for a
gi&en protein0
A. Dialysis
!. Affinity chromatography
". 'el filtration chromatography
D. ;on exchange chromatography
#. #lectrophoresis
The answer is: B
#ach of the techniques listed separates proteins from each other and from other biologic
molecules based upon characteristics such as si6e solubility and charge. 1o$e&er only
affinity chromatography can use the high affinity of proteins for specific chemical groups
or the specificity of immobili6ed antibodies for unique proteins. ;n affinity
chromatography a specific compound that binds to the desired protein4such as an
antibody a polypeptide receptor or a substrate4is co&alently bound to the column
material. A mixture of proteins is added to the column under conditions ideal for binding
the protein desired and the column is then $ashed $ith buffer to remo&e unbound
proteins. The protein is eluted either by adding a high concentration of the original
binding material or by ma*ing the conditions unfa&orable for binding (e.g. changing the
p1). The other techniques are less specific than affinity binding for isolating proteins.
Dialysis separates large proteins from small molecules. ;on exchange chromatography
separates proteins $ith an o&erall charge of one sort from proteins $ith an opposite
charge (e.g. negati&e from positi&e). 'el filtration chromatography separates on the basis
of si6e. #lectrophoresis separates proteins on the principle that net charge influences the
rate of migration in an electric field.
,5.
A solution of glutamic acid is titrated from p1 1.. to B.. by the addition of : m> of a
solution of 1 M Na71. /hat is the approximate number of millimoles of amino acid in
the sample (pKa1 F +.1- pKa+ F ,.+: pKa5 F -.9B)0
A. 1.:
!. 5..
". 9..
D. 1+..
#. 1E..
The answer is: B
To reach p1 B.. approximately 1..N of the %carboxyl group (pKa1 F +.1-) and -.N of
the %carboxyl group (pKa+ F ,.+:) of glutamic acid must be dissociated. At that p1
approximately t$ice the amount of Na71 as glutamic acid molecules has been utili6ed to
titrate the t$o carboxyl groups. )ince each milliliter of a 1 M Na71 solution contains 1
mmol of 71
J
ion about 5 mmol of the amino acid is present.
,,.
/hich one of the follo$ing proteolytic en6ymes is acti&ated by acid hydrolysis of the
proen6yme form0
A. Trypsin
!. "hymotrypsin
". #lastase
D. 3epsin
#. "arboxypeptidase
The answer is: D
3epsin is secreted in a proen6yme form in the stomach. Cnli*e the majority of
proen6ymes it is not acti&ated by protease hydrolysis. ;nstead spontaneous acid
hydrolysis at p1 + or lo$er con&erts pepsinogen to pepsin. 1ydrochloric acid secreted by
the stomach lining creates the acid en&ironment. All the en6ymes secreted by the
pancreas are acti&ated at the same time upon entrance into the duodenum. This is
accomplished by trypsin hydrolysis of the inacti&e proen6ymes trypsinogen
chymotrypsinogen procarboxypeptidase and proelastase. 3rimer amounts of trypsin are
deri&ed from trypsinogen by the action of enteropeptidase secreted by the cells of the
duodenum.
,:.
/hich one of the follo$ing structures may be classified as a hydrophobic amino acid at
p1 B..0
A. ;soleucine
!. Arginine
". Aspartic acid
D. >ysine
#. Threonine
The answer is: A
The carbon next to a carboxyl ("F7) group may be designated as the carbon $ith
subsequent carbons as etc. %amino acids contain an amino group on their
carbon as distinguished from compounds li*e %aminobutyric acid in $hich the amino
group is t$o carbons do$n ( %carbon). ;n %amino acids the amino acid carboxylic acid
and the side chain or 2 group are all bound to the central %carbon $hich is thus
asymmetric (except $hen 2 is hydrogen as for glycine). Amino acids are classified as
acidic neutral hydrophobic neutral hydrophilic or basic depending on the charge or
partial charge on the 2 group at p1 B... 1ydrophobic ($ater%hating) groups are carbon%
hydrogen chains li*e those of leucine isoleucine glycine or &aline. !asic 2 groups such
as those of lysine and arginine carry a positi&e charge at physiologic p1 o$ing to
protonated amide groups $hile acidic 2 groups such as glutamic acid carry a negati&e
charge o$ing to ioni6ed carboxyl groups. Threonine $ith its hydroxyl side chain is
neutral at physiologic p1. Neutral hydrophilic side chains ha&e uncharged but polar or
partially charged groups.
,9.
/hich of the follo$ing amino acids is ioni6able in proteins0
A. >eucine
!. 1istidine
". Daline
D. Alanine
#. 'lycine
The answer is: B
#xcept for terminal amino acids all %amino groups and all %carboxyl groups are
utili6ed in peptide bonds. Thus only amino acids $ith side chains may be considered. 7f
these B of the +. common amino acids ha&e easily ioni6able side chains. These are the
basic amino acids lysine arginine and histidine8 the acidic amino acids aspartate and
glutamate8 and tyrosine and cysteine. >eucine &aline and alanine ha&e hydrocarbon side
chains.
,B.
The oxygen carrier of muscle is the globular protein myoglobin. /hich one of the
follo$ing amino acids is highly li*ely to be locali6ed $ithin the interior of the molecule0
A. Arginine
!. Aspartic acid
". 'lutamic acid
D. Daline
#. >ysine
The answer is: D
The structure of myoglobin is illustrati&e of most $ater%soluble proteins. 'lobular
proteins tend to fold into compact configurations $ith nonpolar cores. The interior of
myoglobin is composed almost exclusi&ely of nonpolar hydrophobic amino acids li*e
&aline leucine phenylalanine and methionine. ;n contrast polar hydrophilic residues
such as arginine aspartic acid glutamic acid and lysine are found mostly on the surface
of the $ater%soluble protein.
,E.
A child stops ma*ing de&elopmental progress at age + years and de&elops coarse facial
features $ith thic* mucous drainage. )*eletal deformities appear o&er the next year and
the child regresses to a &egetati&e state by age 1. years. The childAs urine tests positi&e
for glycosaminoglycans that include $hich of the follo$ing molecules0
A. "ollagen B. %aminobutyric acid C. 1eparan sulfate D. 'lycogen
E. =ibrillin
The answer is: C
'lycosaminoglycans (mucopolysaccharides) are polysaccharide chains that may be
bound to proteins as proteoglycans. #ach proteoglycan is a complex molecule $ith a core
protein that is co&alently bound to glycosaminoglycans4repeating units of disaccharides.
The amino sugars forming the disaccharides contain negati&ely charged sulfate or
carboxylate groups. The primary glycosaminoglycans found in mammals are hyaluronic
acid heparin heparan sulfate chondroitin sulfate and *eratan sulfate. ;nborn errors of
glycosaminoglycan degradation cause neurodegeneration and physical stigmata described
by the outmoded term @gargoylism.@ 'lycogen is a polysaccharide of glucose used for
energy storage and has no sulfate groups. "ollagen and fibrillin are important proteins in
connecti&e tissue. %aminobutyric acid is a %amino acid in&ol&ed in neurotransmission.
,-.
Cnder normal conditions in blood $hich of the follo$ing amino acid residues of albumin
is neutral0
A. Arginine
!. Aspartate
". 'lutamine
D. 'lutamate
#. 1istidine
The answer is: C
;n blood and other solutions at physiologic p1 (approximately B..) only terminal
carboxyl groups terminal amino groups and ioni6able side chains of amino acid residues
in proteins ha&e charges. The basic amino acids lysine arginine and histidine ha&e
positi&e charges (protonated amines). The acidic amino acids aspartate and glutamate
ha&e negati&e charges (ioni6ed carboxyls). 'lutamine possesses an uncharged but
hydrophilic side chain.
:..
/hich of the substances belo$ is primarily found in tendons0
A. "ollagen
!. Troponin
". =ibrillin
D. =ibrin
#. =ibronectin
The answer is: A
"ollagens are insoluble proteins that ha&e great tensile strength. They are the main fibers
composing the connecti&e tissue elements of s*in bone teeth tendons and cartilage.
"ollagen is composed of tropocollagen a triple%stranded helical rod rich in glycine
proline and hydroxyproline residues. Troponin is found in muscle fibrillin in heart
&al&es blood &essels and ligaments <it is defecti&e in (arfanAs syndrome (1:,B..)?.
=ibrin is a component of blood clots and fibronectin is a component of extracellular
matrix.
:1.
During synthesis of mature collagen fiber $hich one of the follo$ing steps $ould occur
$ithin the fibroblast0
A. 1ydrolysis of procollagen to form collagen
!. 'lycosylation of proline residues
". =ormation of a triple helix
D. =ormation of co&alent cross%lin*s bet$een molecules
#. Assembly of the collagen fiber
The answer is: C
The connecti&e tissue fiber collagen is synthesi6ed by fibroblasts. 1o$e&er because the
length of the finished collagen fibers is many times greater than that of the cell of origin
a portion of assembly occurs extracellularly. The intracellular formation of the
biosynthetic precursor of collagen procollagen peptides pro% 1(;) and pro% + occurs in
the follo$ing stepsG (1) synthesis of polypeptides (+) hydroxylation of proline and lysine
residues (5) glycosylation of lysine residues (proline residues are not glycosylated) (,)
formation of the triple helix and (:) secre%tion. 7nce outside the fibroblasts procollagen
molecules are acti&ated by fibroblast%specific procollagen peptidases. !efore specific
proteolytic clea&age of procollagen tropocollagen bundles do not assemble into collagen
fibers. 7nce the collagen fibers are formed aldo cross%lin*s bet$een lysine residues and
histidine%aldo cross%lin*s are formed. These cross%lin*s co&alently bind the collagen
chains to one another. The extent and type of cross%lin*ing determines the flexibility and
strength of the collagen mass formed.
:+.
A 9:%year%old obese male presents $ith se&ere indigestion and chest pain after a spicy
meal. A lactate dehydrogenase (>D1) le&el obtained to e&aluate possible myocardial
infarction is normal but the laboratory recommends that >D1 iso6ymes be performed.
The managing physician *no$s that lactate dehydrogenase is composed of t$o different
polypeptide chains arranged in the form of a tetramer. Assuming that all possible
combinations of the different polypeptide chains occur ho$ many iso6yme forms of
lactate dehydrogenase must be measured0
A. T$o
!. Three
". =our
D. =i&e
#. )ix
The answer is: D
;so6ymes are multiple forms of a gi&en en6yme that occur $ithin a gi&en species. )ince
iso6ymes are composed of different proteins analysis by electrophoretic separation can
be done. >actate dehydrogenase is a tetramer composed of any combination of t$o
different polypeptides 1 and (. Thus the possible combinations are 1, 15(1 1+(+
11(5 and (,. Although each combination is found in most tissues (, predominates in
the li&er and s*eletal muscle $hile 1, is the predominant form in the heart. /hite and
red blood cells as $ell as brain cells contain primarily intermediate forms. The (, forms
of the iso6yme seem to ha&e a higher affinity for pyru&ate compared $ith the 1, form.
=ollo$ing a myocardial infarction the 1, (>D11) type of lactate dehydrogenase rises
and reaches a pea* approximately 59 h later. #le&ated >D11 le&els may signal
myocardial disease e&en $hen the total lactate dehydrogenase le&el is normal.
:5.
/hich of the follo$ing mutations $ould produce a se&ere thalassemia0
A. Deletion of one %globin locus B. Deletion of one %globin locus C.
7xidation of heme groups to produce methemoglobin D. Altered 2NA processing at
both %globin loci E. )ic*le cell anemia
The answer is: D
(utations that alter the balance of % and %globin synthesis from their respecti&e loci on
chromosomes 19 and 11 produce thalassemias. )ince these loci are autosomal mutations
at both homologous loci are required to produce se&ere thalassemia as $ith the
thalassemias that in&ol&ed altered 2NA splicing at both %globin loci. (ethemoglobin is
hemoglobin $ith the iron oxidi6ed from the ferrous (=e
II
) to the ferric (=e
III
) state.
(ethemoglobin cannot bind oxygen so there is a specific en6yme (methemoglobin
reductase) and reducing substances li*e glutathione in red cells that maintain hemoglobin
iron in its reduced state. 3oint mutations that cause amino acid substitutions produce an
abnormal hemoglobin rather than imbalance chain synthesis. )ic*le cell anemia (1,1-..)
is one example in $hich both %globin chains ha&e a &aline replacing glutamine. The
mutant %globin chains in hemoglobin ) ha&e a @stic*y patch@ on their surface that is
particularly adhesi&e $hen hemoglobin ) is deoxygenated. =or this reason indi&iduals
$ith sic*le cell anemia are prone to thrombotic crises (stro*es heart attac*s ischemic
extremities) $hen they become dehydrated (increased hemoglobin concentration) or
hypoxic (more deoxyhemoglobin )). There are also mutant hemoglobins (hemoglobin (
etc.) that predispose to oxidation of the iron group in heme producing higher
concentrations of methemoglobin and cyanosis (bluish color of the lips and fingertips).
:,.
An increased affinity of hemoglobin for 7+ may result from $hich of the follo$ing0
A. ;nitial binding of 7+ to one of the four sites a&ailable in each deoxyhemoglobin
molecule
!. 1igh p1
". 1igh "7+ le&els
D. 1igh +5%bisphosphoglycerate (!3') le&els $ithin erythrocytes
#. Acidosis
The answer is: A
;n addition to its function as a carrier of 7+ and "7+ hemoglobin buffers sudden
additions of acid or base to the blood by &irtue of the histidine 1,9 on each chain.
1o$e&er protonation of the imida6ole of histidine causes deoxygenation of hemoglobin.
Thus decreased binding of 7+ occurs in the high%p1 conditions of acidosis. +5%
bisphosphoglycerate (!3') binds specifically to deoxyhemoglobin8 that is !3' cross%
lin*s positi&ely charged residues on the chain thereby decreasing oxygen affinity and
stabili6ing the deoxygenated form of hemoglobin. The addition of each 7+ molecule to
deoxyhemoglobin requires the brea*age of salt lin*s such as those formed by +5%!3'.
#ach subsequent 7+ molecule requires the brea*age of fe$er salt lin*s. Thus initial 7+
binding actually results in an increased affinity for subsequent 7+ binding $hich in turn
results in a cooperati&e allosteric binding mechanism. "7+ reacts re&ersibly $ith the
amino acid terminals of hemoglobin to create carbaminohemoglobin $hich is negati&ely
charged and $hich forms salt bridges stabili6ing deoxyhemoglobin. 1ence "7+ binding
lo$ers the affinity of hemoglobin for 7+.
::.
The &elocity%substrate cur&e belo$ characteri6es an allosteric en6yme system. The cur&e
demonstrates that
A. A modifier changes the binding constant for the substrate but not the &elocity of
the reaction B. A modifier binding to the allosteric site can also affect the
catalytic site C. !inding of the substrate is independent of its concentration
D. !inding of the modifier is independent of its concentration E. !inding
of substrate to the allosteric site displaces modifier
The answer is: B
/hen a modifier binds at the allosteric site it affects the acti&e site by altering Vmax and
Km. The substrate binds to the acti&e or catalytic site $here it is modified. !inding of
both substrate and modifier is of course concentration%dependent. The &elocity of an
allosteric en6yme reaction depends on the concentration of both the substrate and the
modifier.
:9.
/hich one of the follo$ing en6ymes is regulated primarily through allosteric interaction0
A. "hymotrypsin
!. 3yru&ate dehydrogenase
". 'lycogen phosphorylase
D. 'lycogen synthase
#. Aspartate transcarbamoylase
The answer is: E
Aspartate transcarbamoylase $hich controls the rate of pyrimidine synthesis in
mammals is negati&ely inhibited by the allosteric effector cytidine triphosphate an end
product of pyrimidine synthesis. The allosteric modulation occurs &ia the binding of
effectors at the regulatory site of the en6yme. Nonco&alent bonds are formed during the
binding bet$een effector and en6yme. ;n contrast all the other en6ymes are acti&ated or
deacti&ated by co&alent modification. "hymotrypsinogen is secreted as an inacti&e
proen6yme (6ymogen) in pancreatic juice and is irre&ersibly acti&ated by trypsin clea&age
of a specific peptide bond. 'lycogen phosphorylase is re&ersibly acti&ated by
phosphorylation of a specific serine residue. At the same time glycogen synthase is
re&ersibly deacti&ated by phosphorylation of a specific serine residue thereby pre&enting
a futile cycle of brea*do$n and resynthesis of glycogen. 3yru&ate dehydrogenase also is
re&ersibly inacti&ated by phosphorylation of a specific serine residue. ;n all four
en6ymes a single discrete co&alent modification leads to conformational changes that
allo$ the s$itching on or off of en6yme acti&ity.
:B.
7f the six cur&es labeled in the >ine$ea&er%!ur* graph belo$ three represent the effects
of . mM, : mM, and 1: mM of a competiti&e inhibitor on a hypothetical en6yme. /hich
of the cur&es most li*ely represents the 1:%mM concentration of the competiti&e
inhibitor0
A. A
!. !
". "
D. D
#. #
The answer is: A
"ompetiti&e inhibitors resemble the structure of the substrate and compete $ith the
substrate to bind to the acti&e site of the en6yme. =or this reason Km increases $ith
increasing inhibitor concentration $hile Vmax remains the same. That is Vmax can be
reached at substrate concentrations sufficiently high to o&ercome the inhibitor. )ince the
x axis intercept represents J1MKm and the y axis intercept represents 1MVmax only cur&es A
O and D sho$ changes in Km $ith no changes in Vmax. /hen J1MKm is interpreted
properly the highest Km &alue is gi&en by cur&e A.
:E.
/hich of the follo$ing en6ymes exhibits a hyberbolic cur&e $hen initial reaction
&elocity is plotted against substrate concentration0
A. Aspartate transcarbamoylase
!. 3hosphofructo*inase
". 1exo*inase
D. 3yru&ate *inase
#. >actate dehydrogenase
The answer is: E
Nonregulatory en6ymes such as lactate dehydrogenase typically exhibit a hyperbolic
saturation cur&e $hen initial &elocity is plotted against substrate concentration. #n6ymes
at *ey points in metabolic path$ays are typically allosteric4their &elocities at a gi&en
substrate concentration may be altered due to effects of metabolites in the path$ay.
Allosteric en6ymes typically exhibit sigmoidal *inetics. #xamples of allosteric en6ymes
include aspartate transcarbamoylase $hich is inhibited by cytidine triphosphate ("T3)8
phosphofructo*inase $hich is inhibited by adenosine triphosphate (AT3) and acti&ated
by fructose +9%bisphosphate8 hexo*inase $hich is inhibited by glucose%9%phosphate8 and
pyru&ate *inase $hich is inhibited by AT3. Allosteric en6ymes produce sigmoidal
*inetics $hen substrate concentration is plotted against reaction &elocity. ;n contrast
hyperbolic plots are obser&ed $ith (ichaelis%(enten en6ymes. The binding of effector
molecules such as end products or second messengers to regulatory subunits of
allosteric en6ymes can either positi&ely or negati&ely regulate catalytic subunits.
:-.
/hich of the follo$ing carbohydrates $ould be most abundant in the diet of strict
&egetarians0
A. Amylose
!. >actose
". "ellulose
D. (altose
#. 'lycogen
The answer is: C
"ellulose the most abundant compound *no$n is the structural fiber of plants and
bacterial $alls. ;t is a polysaccharide consisting of chains of glucose residues lin*ed by
1 , bonds. )ince humans do not ha&e intestinal hydrolases that attac* 1 , lin*ages
cellulose cannot be digested but forms an important source of @bul*@ in the diet. >actose
is a disaccharide of glucose and galactose found in mil*. Amylose is an unbranched
polymer of glucose residues in %1, lin*ages. 'lycogen is a branched polymer of
glucose $ith both %1, and %19 lin*ages. (altose is a disaccharide of glucose $hich
is usually the brea*do$n product of amylose.
9..
"hronic alcoholics require more ethanol than do nondrin*ers to become intoxicated
because of a higher le&el of specific en6yme. 1o$e&er independent of specific en6yme
le&els the a&ailability of $hat other substance is rate%limiting in the clearance of ethanol0
A. NAD1
!. NADI
". =AD1
D. =ADI
#. NAD31
The answer is: B
;n humans ethanol is cleared from the body by oxidation cataly6ed by t$o NAD
I
%lin*ed
en6ymesG alcohol dehydrogenase and acetaldehyde dehydrogenase. These en6ymes act
mainly in the li&er to con&ert alcohol to acetaldehyde and acetate respecti&ely. ;n chronic
alcoholics alcohol dehydrogenase may be ele&ated some$hat. The NAD1 le&el is
significantly increased in the li&er during oxidation of alcohol o$ing to the consumption
of NAD
I
. This leads to a s$amping of the normal means of regenerating NAD
I
. Thus
NAD
I
becomes the rate%limiting factor in oxidation of excess alcohol.
91.
/hich one of the follo$ing en6ymes cataly6es high%energy phosphorylation of substrates
during glycolysis0
A. 3yru&ate *inase
!. 3hosphoglycerate *inase
". Triose phosphate isomerase
D. Aldolase
#. 'lyceraldehyde%5%phosphate dehydrogenase
The answer is: E
1igh%energy phosphate bonds are added to the substrates of glycolysis at three steps in
the path$ay. 1exo*inase4or in the case of li&er gluco*inase4adds phosphate from
AT3 to glucose to form glucose%9%phosphate. )trictly spea*ing this is not al$ays
considered a step of the glycolytic path$ay. 3hosphofructo*inase uses AT3 to con&ert
fructose%9%phosphate to fructose%19%phosphate. Csing NAD
I
in an oxidation%reduction
reaction inorganic phosphate is added to glyceraldehyde%5%phosphate by the en6yme
glyceraldehyde%5%phosphate dehydrogenase to form 15%diphosphoglycerate. The
en6ymes phosphoglycerate *inase and pyru&ate *inase transfer substrate high%energy
phosphate groups to AD3 to form AT3.
9+.
/hich one of the follo$ing en6ymes is common to both glycolysis and gluconeogenesis0
A. 3yru&ate *inase
!. 3yru&ate carboxylase
". 1exo*inase
D. 3hosphoglycerate *inase
#. =ructose%19%bisphosphatase
The answer is: D
All the en6ymes listed are specific to either glycolysis or gluconeogenesis except for
phosphoglycerate *inase. ;t is one of se&en en6ymes common to both glycolysis and
gluconeogenesis. The en6ymes hexo*inase phosphofructo*inase and pyru&ate *inase
cataly6e irre&ersible reactions unique to glycolysis. ;n order for gluconeogenesis to occur
the three irre&ersible reactions must be replaced. 3yru&ate is synthesi6ed into
phosphoenolpyru&ate by a t$o%step reaction. =irst oxaloacetate is formed by
carboxylation in the presence of pyru&ate carboxylase. Then phosphoenolpyru&ate
carboxy*inase decarboxylates and phosphorylates oxaloacetate in the presence of 'T3.
The next irre&ersible step to be bypassed in gluconeogenesis requires fructose%9%
phosphate to be produced by the action of fructose%19%phosphatase on fructose%19%
phosphate. /hen glucose%9%phosphate is finally produced during gluconeogenesis it is
con&erted to glucose by glucose%9%phosphatase an en6yme unique to the endoplasmic
reticulum. The free glucose may then diffuse from the li&er into the bloodstream. 7f the
en6ymes gi&en as possible ans$ers only phosphoglycerate *inase cataly6es a re&ersible
reaction common to both glycolysis and gluconeogenesis.
;n&ol&ment of the citric acid cycle in transamination and gluconeogenesis. !old arro$s
indicate the main path$ay of gluconeogenesis.
(Reproduced, with permission, from Murray RK, ranner !K, Mayes "#, Rodwell V$%
1arperAs !iochemistry &'(e) *ew Yor+, Mcraw-,ill, &---% ./0)1
23)
During the first $ee* of a diet of 1:.. calories per day the oxidation of glucose &ia
glycolysis in the li&er of a normal :-%*g (15.%lb) $oman is inhibited by the lo$ering of
$hich one of the follo$ing0
A. "itrate
!. AT3
". =atty acyl "oA
D. Qetone bodies
#. =ructose%+9%bisphosphate
The answer is: E
The main control of glycolysis is through the en6yme phosphofructo*inase. This en6yme
is controlled by a high le&el of AT3 $hich inhibits it or a high le&el of fructose%+9%
bisphosphate (=%+9%!3) $hich acti&ates it. The inhibitory effect of AT3 is potentiated by
citrate $hile high A(3 le&els re&erse it. During fasting $hen blood glucose le&els are
lo$ a glucagon%signaled increase of li&er cyclic A(3 leads to the acti&ation of a
phosphatase that hydroly6es the +%phosphoryl group from =%+9%!3. The same glucagon%
stimulated cascade deacti&ates the *inase that phosphorylates fructose%9%phosphate. The
subsequent lo$ering of =%+9%!3 inacti&ates phosphofructo*inase.
9,
/hich one of the follo$ing en6ymes cataly6es phosphorylation $ith the use of inorganic
phosphate0
A. 1exo*inase
!. 3hosphofructo*inase
". 'lyceraldehyde%5%phosphate dehydrogenase
D. 3hosphoglycerate *inase
#. 3yru&ate *inase
The answer is: C
All the en6ymes named are glycolytic en6ymes that carry out phosphorylation of glucose%
deri&ed substrates or of AD3 to form AT3. 1o$e&er only the reaction cataly6ed by
glyceraldehyde%5%phosphate dehydrogenase is a phosphorylation reaction coupled to
oxidation that uses inorganic phosphate. ;n this reaction glyceraldehyde%5%phosphate is
con&erted to 15%bisphosphoglycerate by the addition of inorganic phosphate and the
oxidation of glyceraldehyde%5%phosphate $ith the concomitant reduction of NAD
I
to
NAD1 I 1
I
. This reaction is an example of a high%energy phosphate compound being
produced by an oxidation%reduction reaction. The oxidation of the aldehyde group at "1
of glyceraldehyde%5%phosphate pro&ides the energy for the reaction. The 15%
bisphosphoglycerate can then be utili6ed to phosphorylate AD3 to AT3 through the action
of phosphoglycerate *inase $hich is the next step in the glycolytic path$ay.
9:.
After a $ell%rounded brea*fast $hich of the follo$ing $ould be expected to occur0
A. ;ncreased acti&ity of pyru&ate carboxylase
!. Decreased acti&ity of acetyl "oA carboxylase
". Decreased rate of glycogenolysis
D. Decreased rate of protein synthesis
#. ;ncreased acti&ity of phosphoenolpyru&ate carboxy*inase
The answer is: C
During an o&ernight fast glycogenolysis and gluconeogenesis occur to some degree.
=ollo$ing a $ell%rounded brea*fast amino acids are a&ailable for protein synthesis
glycogen synthesis occurs from excess glucose gluconeogenesis decreases and fatty acid
synthesis occurs from excess acetyl "oA produced from dietary sources. Thus the
acti&ity of en6ymes of gluconeogenesis (pyru&ate carboxylase and phosphoenolpyru&ate
carboxy*inase) decreases $hile the acti&ity of en6ymes of fatty acid synthesis (acetyl
"oA carboxylase) increases.
99.
/hich of the follo$ing hormones stimulates gluconeogenesis0
A. 3rogesterone
!. 'lucagon
". Aldosterone
D. #pinephrine
#. Thyroxine
=. 'ro$th hormone
'. ;nsulin
1. 'lucocorticoids
The answer is: B
The balance and integration of the metabolism of fats and carbohydrates are mediated by
the hormones insulin glucagon epinephrine and norepinephrine. All of these hormones
exercise acute effects upon metabolism. 'lucagon stimulates gluconeogenesis and bloc*s
glycolysis. /hen blood sugar le&els get lo$ the cells of the pancreas release glucagon.
The main targets of glucagon are the li&er and adipose tissue. ;n the li&er glucagon
stimulates the cyclic A(3Jmediated cascade that causes phosphorylation of
phosphorylase and glycogen synthesis. This effecti&ely turns off glycogen synthase and
turns on glycogen phosphorylase thereby causing a brea*do$n of glycogen and a
production of glucose in li&er $hich ultimately raises blood glucose le&els. ;nsulin and
glucagon are t$o antagonistic hormones that maintain the balance of sugar and fatty
acids in blood. ;nsulin is produced by the cells of the pancreas and its release is
stimulated by high le&els of glucose in the blood. ;t has a number of effects but its major
effect is to allo$ the entry of glucose into cells. ;nsulin also allo$s the dephosphorylation
of *ey regulatory en6ymes. The consequence of these actions is to allo$ glycogen
synthesis and storage in both muscle and li&er suppression of gluconeogenesis
acceleration of glycolysis promotion of the synthesis of fatty acids and promotion of the
upta*e and synthesis of amino acids into protein. All in all insulin acts to promote
anabolism.
9B.
A Nigerian medical student studying in the Cnited )tates de&elops hemolytic anemia
after ta*ing the oxidi6ing antimalarial drug pamaquine. This se&ere reaction is most li*ely
due to
A. 'lucose%9%phosphate dehydrogenase deficiency
!. "oncomitant scur&y
". Ditamin " deficiency
D. Diabetes
#. 'lycogen phosphorylase
The answer is: A
7ne of the $orldAs most common en6yme deficiencies is glucose%9%phosphate%
dehydrogenase deficiency (5.:-..). This deficiency in erythrocytes is particularly
pre&alent among African and (editerranean males. A deficiency in glucose%9%phosphate
dehydrogenase bloc*s the pentose phosphate path$ay and NAD31 production. /ithout
NAD31 to maintain glutathione in its reduced form erythrocytes ha&e no protection
from oxidi6ing agents. This O%lin*ed recessi&e deficiency is often diagnosed $hen
patients de&elop hemolytic anemia after recei&ing oxidi6ing drugs such as pamaquine or
after eating oxidi6ing substances such as fa&a beans.
9E.
Among the many molecules of high%energy phosphate compounds formed as a result of
the functioning of the citric acid cycle one molecule is synthesi6ed at the substrate le&el.
;n $hich of the follo$ing reactions does this occur0
A. "itrate %*etoglutarate B. %*etoglutarate succinate C. )uccinate
fumarate D. =umarate malate E. (alate oxaloacetate
The answer is: B
A molecule of guanosine triphosphate is synthesi6ed from guanosine diphosphate and
phosphate at the cost of hydroly6ing succinyl "oA to succinate and "oA. This constitutes
substrate%le&el phosphorylation and in contrast to oxidati&e phosphorylation this is the
only reaction in the citric acid cycle that directly yields a high%energy phosphate bond.
The sequence of reactions from %*etoglutarate to succinate is cataly6ed by the %
*etoglutarate dehydrogenase complex and succinyl%"oA synthetase respecti&ely.
%*etoglutarate I NAD
I
I acetyl "oA succinyl "oA I "7+ I NAD1
succinyl "oA I 3i I 'D3 succinate I 'T3 I acetyl "oA
9-.
2eduction of $hich one of the follo$ing substrates leads to a reducing equi&alent in a
step of the citric acid cycle0
A. )uccinyl "oA
!. (alate
". =umarate
D. 7xaloacetate
#. "itrate
The answer is: B
2educing equi&alents are produced at four sites in the citric acid cycle. NAD1 is
produced by the isocitrate dehydrogenaseJcataly6ed con&ersion of %*etoglutarate to
succinyl "oA and by the malate dehydrogenaseJcataly6ed con&ersion of malate to
oxaloacetate. =AD1+ is produced by the succinate dehydrogenaseJcataly6ed con&ersion
of succinate to fumarate. )uccinyl "oA synthetase cataly6es the formation of succinate
from succinyl "oA $ith the concomitant phosphorylation of 'D3 to 'T3.
B..
A child has ingested cyanide from her parentsA garage and is rushed to the emergency
room. /hich of the follo$ing components of the citric acid cycle $ill be depleted first in
this child0
A. NADI cofactor
!. "itrate synthase
". Aconitase
D. "itrate production
#. Acetyl coen6yme A ("oA) production
The answer is: A
"yanide bloc*s respiration by displacing oxygen from hemoglobin. 7xidati&e
phosphorylation in the mitochondria cannot proceed because cyanide cannot oxidi6e
(remo&e electrons) from reduced cofactors li*e NAD1. The citric acid cycle is the major
path$ay for generating AT3 and reducing equi&alents (NAD1 1
I
) from catabolism of
carbohydrates amino acids and lipids. ;nability to regenerate NAD
I
from NAD1
through mitochondrial oxidati&e phosphorylation depletes the cell of NAD
I
and inhibits
the citric acid cycle. =ailure to generate AT3 by oxidati&e phosphorylation using NAD1
from the citric acid cycle depletes the cell of energy and leads to cell and tissue death
(organ failure). #n6ymes (citrate synthase aconitase) and intermediates of the citric acid
cycle (citrate acetyl coen6yme A) need only be present in trace amounts because they are
not consumed.
B1.
A child presents $ith lo$ blood glucose (hypoglycemia) enlarged li&er (hepatomegaly)
and excess fat deposition in the chee*s (cherubic facies). A li&er biopsy re&eals excess
glycogen in hepatocytes. Deficiency of $hich of the follo$ing en6ymes might explain
this phenotype0
A. %11%glucosidase
!. %11%galactosidase
". %1,%glucosidase
D. %1,%galactosidase
#. %19%galactosidase
The answer is: C
The child has symptoms of glycogen storage disease. 'lycogen is a glucose polymer $ith
linear regions lin*ed through the "1 aldehyde of one glucose to the ", alcohol of the
next ( %1,%glucoside lin*age). There are also branches from the linear glycogen polymer
that ha&e %19%glucoside lin*ages. 'lycogen is synthesi6ed during times of carbohydrate
and energy surplus but must be degraded during fasting to pro&ide energy. )eparate
en6ymes for brea*do$n include phosphylases ( %1,%glucosidases) that clea&e linear
regions of glycogen and debranching en6ymes ( %19%glucosidases) that clea&e branch
points. 'lucose%9%phosphatase is needed in the li&er to liberate free glucose from
glucose%9%phosphate pro&iding fuel for other organs. There is no glucose%9%phosphatase
in muscle and muscle glycogenolysis pro&ides energy just for muscle $ith production of
lactate. Deficiencies of more than eight en6ymes in&ol&ed in glycogenolysis including
those mentioned can produce glycogen storage disease.
B+.
A man goes on a hunger stri*e and confines himself to a liquid diet $ith minimal
calories. /hich of the follo$ing $ould occur after , to : h0
A. Decreased cyclic A(3 and increased li&er glycogen synthesis
!. ;ncreased cyclic A(3 and increased li&er glycogenolysis
". Decreased epinephrine le&els and increased li&er glycogenolysis
D. ;ncreased "aII in muscle and decreased glycogenolysis
#. Decreased "aII in muscle and decreased glycogenolysis
The answer is: B
;n the presence of lo$ blood glucose epinephrine or norepinephrine interacts $ith
specific receptors to stimulate adenylate cyclase production of cyclic A(3. "yclic A(3
acti&ates protein *inase $hich cataly6es phosphorylation and acti&ation of phosphorylase
*inase. Acti&ated phosphorylase *inase acti&ates glycogen phosphorylase $hich
cataly6es the brea*do$n of glycogen. 3hosphorylase *inase can be acti&ated in t$o $ays.
3hosphorylation leads to complete acti&ation of phosphorylase *inase. Alternati&ely in
muscle the transient increases in le&els of "a
II
associated $ith contraction lead to a
partial acti&ation of phosphorylase *inase. "a
II
binds to calmodulin $hich is a subunit of
phosphorylase *inase. "almodulin regulates many en6ymes in mammalian cells through
"a
II
binding.
B5.
(cArdleAs disease causes muscle cramps and muscle fatigue $ith increased muscle
glycogen. /hich of the follo$ing en6ymes is deficient0
A. 1epatic hexo*inase
!. (uscle glycogen synthetase
". (uscle phosphorylase
D. (uscle hexo*inase
#. (uscle debranching en6yme
The answer is: C
(uscle phosphorylase deficiency leads to a glycogen storage disease <(cArdleAs disease
(+5+9..)? and in young adults an inability to do strenuous physical $or* because of
muscular cramps resulting from ischemia. The compromised phosphorylation of muscle
glycogen characteristic of (cArdleAs disease compels the muscles to rely on auxiliary
energy sources such as free fatty acids and ambient glucose
B,.
Transfer of 1
I
Me
J
pairs to electron transport carriers decarboxylation and substrate%le&el
phosphorylation occur at some of the steps sho$n in the follo$ing diagram of the citric
acid cycle. All three of these e&ents occur at $hich step0
A. )tep A
!. )tep !
". )tep "
D. )tep D
#. )tep #
The answer is: C
;n the citric acid cycle the con&ersion of %*etoglutarate to succinate results in
decarboxylation transfer of an 1
I
Me
J
pair to NAD1 I 1
I
and the substrate%le&el
phosphorylation of 'D3 to 'T3. The series of reactions in&ol&ed is quite complex. =irst
%*etoglutarate reacts $ith NAD
I
I "oA to yield succinyl "oA I "7+ I NAD1 I 1
I
.
These reactions occur by the catalysis of the %*etoglutarate dehydrogenase complex
$hich contains lipoamide =AD
I
and thiamine pyrophosphate as prosthetic groups.
Cnder the action of succinyl "oA synthetase succinyl "oA cataly6es the phosphorylation
of 'D3 $ith inorganic phosphate coupled to the clea&age of the thioester bond of
succinyl "oA. Thus the production of succinate from %*etoglutarate yields one
substrate%le&el phosphorylation and the production of three AT3 equi&alents from NAD1
&ia oxidati&e phosphorylation.
B:.
A comatose laboratory technician is rushed into the emergency room. )he dies $hile you
are examining her. 1er most dramatic symptom is that her body is literally hot to your
touch indicating an extremely high fe&er. Rou learn that her lab has been $or*ing on
metabolic inhibitors and that there is a high li*elihood that she accidentally ingested one.
/hich one of the follo$ing is the most li*ely culprit0
A. !arbiturates
!. 3iericidin A
". Dimercaprol
D. Dinitrophenol
#. "yanide
The answer is: D
All of the poisons sho$n affect either electron transport or oxidati&e phosphorylation.
Dinitrophenol is unique in that it disconnects the ordinarily tight coupling of electron
transport and phosphorylation. ;n its presence electron transport continues normally $ith
no oxidati&e phosphorylation occurring. ;nstead heat energy is generated. The same
principle is utili6ed in a $ell%controlled $ay by bro$n fat to generate heat in ne$born
humans and cold%adapted mammals. The biological uncoupler in bro$n fat is a protein
called thermogenin. !arbiturates the antibiotic piericidin A the fish poison rotenone
dimercaprol and cyanide all act by inhibiting the electron transport chain at some point.
B9.
As electrons are recei&ed and passed do$n the transport chain sho$n belo$ the &arious
carriers are first reduced $ith acceptance of the electron and then oxidi6ed $ith loss of
the electron. A patient poisoned by $hich of the follo$ing compounds has the most
highly reduced state of most of the respiratory chain carriers0
A. Antimycin A
!. 2otenone
". "arbon monoxide
D. 3uromycin
#. "hloramphenicol
The answer is: C
The electron transport chain sho$n contains three proton pumps lin*ed by t$o mobile
electron carriers. At each of these three sites (NAD1JS reductase cytochrome reductase
and cytochrome oxidase) the transfer of electrons do$n the chain po$ers the pumping of
protons across the inner mitochondrial membrane. The bloc*age of electron transfers by
specific point inhibitors leads to a buildup of highly reduced carriers behind the bloc*
because of the inability to transfer electrons across the bloc*. ;n the scheme sho$n
rotenone bloc*s step A antimycin A bloc*s step ! and carbon monoxide (as $ell as
cyanide and a6ide) bloc*s step #. Therefore a carbon monoxide inhibition leads to a
highly reduced state of all of the carriers of the chain. 3uromycin and chloramphenicol
are inhibitors of protein synthesis and ha&e no direct effect upon the electron transport
chain.
BB.
/hich of the follo$ing compounds is a member of the electron transport chain0
A. 7ctanoyl carnitine
!. "ytochrome c
". 2educed nicotinamide adenine dinucleotide (NAD1)
D. 3almitoyl carnitine
#. "arnitine
The answer is: C
NAD1 is generated by many biochemical reactions and used as a reducing agent for
biosynthesis. ;n addition it is part of the transport chain that con&eys electrons to oxygen
in the mitochondria. "arnitine binds to fatty acids such as octanoic ("E) or palmitoic
("19) acid forming acylcarnitines that can be transported into the mitochondria for
oxidation. A patient $ith systemic deficiency of carnitine cannot utili6e fatty acids for
production of energy. All tissues become glucose%dependent resulting in hypoglycemia
and potential death during o&ernight fasting.
BE.
'i&en that the standard free energy change ( 'TA) for the hydrolysis of AT3 is JB.5
*calMmol and that for the hydrolysis of glucose%9%phosphate is J5.5 *calMmol $hat is the
'TA for the phosphorylation of glucose0
glucose I AT3 glucose%9%phosphate I AD3
A. J1..9 *calMmol
!. JB.5 *calMmol
". J,.. *calMmol
D. I,.. *calMmol
#. I1..9 *calMmol
The answer is: C
'luco*inase in the li&er or hexo*inase in other tissues cataly6es the phosphorylation of
glucose as the first step of glycolysis. The equilibrium lies far to the right for the reaction
as $rittenG
glucose I AT3 glucose%9%phosphate I AD3 I 3i
The 'TA for the hydrolysis of glucose%9%phosphate is J5.5 *calMmol. Thus the 'TA of
the re&erse reaction is I5.5 *calMmol. )ince the 'TA for the hydrolysis of AT3 is JB.5
*calMmol the 'TA for the reaction isG
(JB.5 *calMmol) I (I5.5 *calMmol) F J,.. *calMmol
The phosphorylation of glucose is a thermodynamically fa&orable reaction.
B-.
/hich of the follo$ing reactions generates AT30
A. 'lucose%9%phosphate to fructose%9%phosphate
!. 'lucose to glucose%9%phosphate
". =ructose%9%phosphate to fructose%19%diphosphate
D. 3hosphoenolpyru&ate to pyru&ate
#. 3yru&ate to lactate
The answer is: D
AT3 is synthesi6ed by t$o reactions in glycolysis. The first molecule of AT3 is generated
by phosphoglycerate *inase con&erting 15%diphosphoglycerate to 5%phosphoglycerate.
The second molecule of AT3 is generated by pyru&ate *inase con&erting
phosphoenolpyru&ate to pyru&ate.
E..
/hich one of the follo$ing en6ymes cataly6es high%energy phosphorylation of substrates
during glycolysis0
A. 3yru&ate *inase
!. 3hosphoglycerate *inase
". Triose phosphate isomerase
D. Aldolase
#. 'lyceraldehyde%5%phosphate dehydrogenase
The answer is: E
1igh%energy phosphate bonds are added to the substrates of glycolysis at three steps in
the path$ay. 1exo*inase4or in the case of the li&er gluco*inase4adds phosphate from
AT3 to glucose to form glucose%9%phosphate. )trictly spea*ing this is not al$ays
considered a step of the glycolytic path$ay. 3hosphofructo*inase uses AT3 to con&ert
fructose%9%phosphate to fructose%19%phosphate. Csing NAD
I
in an oxidation%reduction
reaction inorganic phosphate is added to glyceraldehyde%5%phosphate by the en6yme
glyceraldehyde%5%phosphate dehydrogenase to form 15%diphosphoglycerate. The
en6ymes phosphoglycerate *inase and pyru&ate *inase transfer substrate high%energy
phosphate groups to AD3 to form AT3.
E1.
/hich reaction in the follo$ing figure occurs in both muscle and li&er but has
substantially different qualities in the t$o0
A. 2eaction A
!. 2eaction !
". 2eaction "
D. 2eaction D
#. 2eaction #
The answer is: B
The con&ersion of glucose to glucose%9%phosphate is different in li&er and muscle. ;n
muscle and most other tissues hexo*inase regulates the con&ersion of glucose to glucose%
9%phosphate. /hen the major regulatory en6yme of glycolysis phosphofructose *inase is
turned off the le&el of fructose%9%phosphate increases and in turn the le&el of glucose%9%
phosphate rises because it is in equilibrium $ith fructose%9%phosphate. 1exo*inase is
inhibited by glucose%9%phosphate. 1o$e&er in the li&er glucose is phosphorylated e&en
$hen glucose%9%phosphate le&els are high because the en6yme regulating transformation
of glucose into glucose%9%phosphate is gluco*inase. 'luco*inase is not inhibited by
glucose%9%phosphate in the li&er. /hile hexo*inase has a lo$ Km for glucose and is
capable of acting upon lo$ le&els of blood glucose gluco*inase has a high Km for glucose
and is effecti&e only $hen glucose is abundant. Therefore $hen blood glucose le&els are
lo$ muscle brain and other tissues are capable of ta*ing up and phosphorylating
glucose $hile the li&er is not. /hen blood glucose is abundant gluco*inase in the li&er
phosphorylates glucose and pro&ides glucose%9%phosphate for the synthesis and storage of
glucose as glycogen.
E+.
A teenage girl is brought to the medical center because of her complaints that she gets too
tired $hen as*ed to participate in gym class. A consulting neurologist finds muscle
$ea*ness in the girlAs arms and legs. /hen no ob&ious diagnosis can be made biopsies of
her muscles are ta*en for tests. "hemistries re&eal greatly ele&ated amounts of
triacylglycerides esterified $ith primarily long%chain fatty acids. 3athology reports the
presence of significant numbers of lipid &acuoles in the muscle biopsy. /hich one of the
follo$ing is the most li*ely diagnosis0
A. =atty acid synthase deficiency
!. Tay%)achs disease
". "arnitine deficiency
D. !iotin deficiency
#. >ipoprotein lipase deficiency
The answer is: C
The most li*ely cause of the symptoms obser&ed is carnitine deficiency. Cnder normal
circumstances long%chain fatty acids coming into muscle cells are acti&ated as acyl
coen6yme A and transported as acyl carnitine across the inner mitochondrial membrane
into the matrix. A deficiency in carnitine $hich is normally synthesi6ed in the li&er can
be genetic8 but it is also obser&ed in preterm babies $ith li&er problems and dialysis
patients. !loc*age of the transport of long%chain fatty acids into mitochondria not only
depri&es the patient of energy production but also disrupts the structure of the muscle
cell $ith the accumulation of lipid droplets. 7ral dietary supplementation usually can
effect a cure. Deficiencies in the carnitine acyltransferase en6ymes ; and ;; can cause
similar symptoms.
E5.
The problem of regenerating NAD
I
from NAD1 for cytoplasmic processes by using
mitochondria is sol&ed in the most energy%efficient manner by $hich one of the follo$ing
intercellular shuttle systems0
A. "itrate pyru&ate shuttle
!. Dihydroxyacetone phosphate %glycerophosphate shuttle
". (alate aspartate shuttle
D. "itrate citrate shuttle
#. >actate pyru&ate shuttle
The answer is: C
NAD1 generated from glycolysis must be relie&ed of an electron to form nicotinamide
adenine dinucleotide (NAD
I
) so that glycolysis may continue. 1o$e&er mitochondrial
membranes are impermeable to both NAD1 and NAD
I
. The solution to this problem is
the transfer of electrons from NAD1 to molecules that tra&erse the membrane. ;n the
glycerophosphate shuttle dihydroxyacetone phosphate (D1A3) is reduced to glycerol%5%
phosphate and thereby regenerates NAD
I
. The glycerol%5%phosphate diffuses into
mitochondria and is oxidi6ed by fla&in adenine dinucleotide (=AD) bac* to D1A3 $hich
can diffuse bac* into the cytosol. The mitochondrial reduced form of fla&in adenine
dinucleotide (=AD1+) produced yields + AT3 in the electron transport chain. ;n the heart
and li&er the more energy%efficient malate%aspartate shuttle mo&es electrons into
mitochondria. "ytoplasmic oxaloacetate is reduced to malate $hich diffuses into the
mitochondria and is oxidi6ed by NADI bac* to oxaloacetate. The mitochondrial NAD1
produced yields 5 AT3 on electron transport. The mitochondrial oxaloacetate is con&erted
to aspartate $hich diffuses into the cytosol $here it is con&erted bac* into cytoplasmic
oxaloacetate.
E,.
/hich one of the follo$ing tissues can metaboli6e glucose fatty acids and *etone bodies
for AT3 production0
A. >i&er
!. (uscle
". 1epatocytes
D. !rain
#. 2ed blood cells
The answer is: B
(uscle cells are the only cells listed that are capable of utili6ing all the energy sources
a&ailable4glucose fatty acids and during fasting *etone bodies. (itochondria are
required for metabolism of fatty acids and *etone bodies. )ince red blood cells
(erythrocytes) do not contain mitochondria no utili6ation of these energy sources is
possible. Although the brain may utili6e glucose and *etone bodies fatty acids cannot
cross the blood%brain barrier. 1epatocytes (li&er cells) are the sites of *etone body
production but the mitochondrial en6yme necessary for utili6ation of *etone bodies is not
present in hepatocytes.
E:.
A child $ith a large head multiple fractures and blue scleras ($hites of the eyes) is
e&aluated for osteogenesis imperfecta (199+..). 7ne study in&ol&es labeling of collagen
chains in tissue culture to assess their mobility by gel electrophoresis. Amino acids
labeled $ith radioacti&e carbon 1, are added to the culture dishes in order to label the
collagen. /hich of the follo$ing amino acids $ould not result in labeled collagen0
A. )erine
!. 'lycine
". Aspartate
D. 'lutamate
#. 1ydroxylserine
The answer is: E
"ollagen has an unusual amino acid composition in that approximately one%third of
collagen molecules are glycine. The amino acid proline is also present in a much greater
amount than in other proteins. ;n addition t$o some$hat unusual amino acids ,%
hydroxyproline and :%hydroxylysine are found in collagen. 1ydroxyproline and
hydroxylysine per se are not incorporated during the synthesis of collagen. 3roline and
lysine are hydroxylated by specific hydroxylases after collagen is synthesi6ed. A reducing
agent such as ascorbate (&itamin ") is needed for the hydroxylation reaction to occur. ;n
its absence the disease *no$n as scur&y occurs. 7nly proline or lysine residues located
on the amino side of glycine residues are hydroxylated. The hydroxylysine residues of
collagen are important sites of glycosylation of disaccharides of glucose and galactose.
E9.
A ne$born becomes progressi&ely lethargic after feeding and increases his respiratory
rate. 1e becomes &irtually comatose responding only to painful stimuli and exhibits
mild respiratory al*alosis. )uspicion of a urea cycle disorder is aroused and e&aluation of
serum amino acid le&els is initiated. ;n the presence of hyperammonemia production of
$hich of the follo$ing amino acids is al$ays increased0
A. 'lycine
!. Arginine
". 3roline
D. 1istidine
#. 'lutamine
The answer is: E
3artial bloc*age of the urea cycle leads to conditions ranging from lethargy and episodic
&omiting to mental retardation. A complete bloc* is incompatible $ith life. A major
reason for the toxicity is the se&ere depletion of AT3 le&els caused by the siphoning off of
%*etoglutarate from the citric acid cycle in an attempt to consume ammonia. 'lutamate
dehydrogenase and glutamine synthetase respecti&ely cataly6e the follo$ing reactionG
%*etoglutarate I N1,
I
glutamate I N1,
I
glutamine
As can be seen this is the re&erse order of steps $hereby glutamine is successi&ely
deaminated first to glutamate and then to %*etoglutarate by the en6ymes glutaminase
and glutamate dehydrogenase respecti&ely. ;t is thought that the high le&el of ammonia
ions shifts the equilibrium of the dehydrogenase in fa&or of the formation of glutamate.
Depending on the step in the urea cycle that is bloc*ed le&els of arginine may be
decreased.
EB.
/hich of the follo$ing metabolites is a precursor of tyrosine0
A. >%dihydroxyphenylalanine (dopa)
!. Dopamine
". Norepinephrine
D. #pinephrine
#. 3henylalanine
The answer is: E
;n humans tyrosine can be formed by the hydroxylation of phenylalanine. This reaction
is cataly6ed by the en6yme phenylalanine hydroxylase. A deficiency of phenylalanine
hydroxylase results in the disease called phenyl*etonuria <3QC(+919..)?. ;n this disease
it is usually the accumulation of phenylalanine and its metabolites rather than the lac* of
tyrosine that is the cause of the se&ere mental retardation ultimately seen. 7nce formed
tyrosine is the precursor of many important signal molecules. "ataly6ed by tyrosine
hydroxylase tyrosine is hydroxylated to form >%dihydroxyphenylalanine (dopa) $hich in
turn is decarboxylated to form dopamine in the presence of dopa decarboxylase. Then
norepinephrine and finally epinephrine are formed from dopamine. All of these are signal
molecules to some degree. Dopa and inhibitors of dopa decarboxylase are used in the
treatment of 3ar*insonAs disease a neurologic disorder. Norepinephrine is a transmitter at
smooth%muscle junctions inner&ated by sympathetic ner&e fibers. #pinephrine and
dopamine are catecholamine transmitters synthesi6ed in sympathetic ner&e terminals and
in the adrenal gland. Tyrosine is also the precursor of thyroxine the major thyroid
hormone and melanin a s*in pigment.
EE.
/hich of the follo$ing amino acids is a precursor to cysteine0
A. Threonine
!. (ethionine
". 'lutamine
D. >ysine
#. Alanine
The ans$er isG !
(ethionine is the only sulfur%containing amino acid of the group and is a precursor for
the sulfur%containing amino acid cysteine. Threonine has an aliphatic side chain $ith a
hydroxyl group glutamine and lysine ha&e amino group side chains and alanine has a
methyl side chain. 1omocysteine is a sulfur%containing amino acid that is not found in
proteins being an intermediate in the formation of cysteine from the sulfur%containing
amino acid methionine. 1omocysteine and methionine are also components of the
acti&ated methyl cycle in $hich )%adenosylmethionine is regenerated. )%
adenosylmethionine is one of the major donors of methyl groups. (ethionine is an
essential amino acid and must be deri&ed from the diet.
E-.
/hich of the follo$ing clinical laboratory obser&ations is suggesti&e of 1artnup disease
(neutral amino acid transport deficiency)0
A. !urnt%sugar smell in urine
!. 1igh plasma phenylalanine le&els
". #xtremely high le&els of citrulline in urine
D. #le&ation of glutamine in blood and urine
#. #le&ated plasma tyrosine and methionine le&els
=. Dar* urine
'. 1igh fecal le&els of tryptophan and indole deri&ati&es
The answer is: G
;n 1artnup disease a defect in the transport process for neutral amino acids is most
pronounced in intestinal and renal transport. Neutral aminoaciduria is obser&ed as $ell as
increased fecal excretion of indole deri&ations due to bacterial con&ersion of unabsorbed
dietary tryptophan. 3ellagra%li*e symptoms can be seen due to the lac* of tryptophan for
niacin biosynthesis. 1yperammonia can be caused by a &ariety of urea cycle defects
including carbamoyl phosphate synthase ; deficiency (+5B5..). The high le&els of N1,
I

in the blood lead to &ery high le&els of glutamine synthesis that may $ell be responsible
for the subsequent brain damage. Deficiency of li&er phenylalanine hydroxylase causes
phenyl*etonuria <3QC(+919..)?. "onsequently high le&els of phenylalanine are not
con&erted to tyrosine. 3henylalanine and its metabolites accumulate in blood leading to
mental retardation if infants are not placed on a phenylalanine%restricted diet. Crine
dar*ens upon standing due to the high le&els of homogentisate that result from of a
deficiency in homogentisate oxidase in the disease al*aptonuria (+.5:..). This is a
deficiency in the path$ay of tyrosine brea*do$n.
-..
/hich of the follo$ing porphyrins gi&es stools their characteristic bro$n color0
A. !ili&erdin
!. Crobilinogen
". 1eme
D. )tercobilin
#. Crobilin
The answer is: D
7nce bile is excreted into the gut bilirubin diglucuronide is hydroly6ed and reduced by
bacteria to form urobilinogen $hich is colorless. (uch of the urobilinogen of the stools
is further oxidi6ed by intestinal bacteria to stercobilin $hich gi&es stools their
characteristic bro$n color. )ome urobilinogen is reabsorbed by the gut into the portal
blood transported to the *idney and con&erted and excreted as urobilin $hich gi&es
urine its characteristic yello$ color.
-1.
/hich of the follo$ing is most characteristic of a sphingolipidosis0
A. (ultifactorial inheritance
!. Dariable acti&ities of abnormal en6yme in different patient tissues
". Deficiency of a hydrolytic en6yme
D. Abnormalities of sphingolipid synthesis
#. Accumulation of ceramide%containing lipids
The answer is: E
)phingolipidoses are lipid storage diseases that exhibit (endelian (autosomal or O%lin*ed
recessi&e) inheritance. To date all lipidoses studied demonstrate accumulation of a
ceramide%containing sphingolipid due to the genetic deficiency of a specific hydrolytic
en6yme in&ol&ed in the brea*do$n of the sphingolipid in question. This leads to the
accumulation of the sphingolipid because its synthetic rate is normal. )ince the decrease
in acti&ity of the abnormal hydrolytic en6yme is similar in all tissues diagnostic tests
measuring the en6yme can easily be set up using s*in biopsies or blood cell
measurements. 1etero6ygous carriers can be screened and recei&e genetic counseling.
#xamples of sphingolipidoses include Tay%)achs disease (+B+E..) )andhoffAs disease
(+9EE..) and 'aucherAs disease (+5.E..).
!iosynthesis of me&alonate sho$ing the critical step of 1('%"oA reductase that is
inhibited by statins. The open and solid circles indicate the fates of carbons in acetyl
"oA.
4&)
;t has been noted that infants placed on extremely lo$%fat diets for a &ariety of reasons
often de&elop s*in problems and other symptoms. This is most often due to
A. >actose intolerance
!. 'lycogen storage diseases
". Antibody abnormalities
D. Deficiency of fatty acid desaturase greater than -
#. Deficiency of chylomicron and D>D> production
The answer is: D
;nfants placed on chronic lo$%fat formula diets often de&elop s*in problems impaired
lipid transport and e&entually poor gro$th. This can be o&ercome by including linoleic
acid to ma*e up 1 to +N of the total caloric requirement. #ssential fatty acids are required
because humans ha&e only
,

:

9
and
-
fatty acid desaturase. 7nly plants ha&e
desaturase greater than
-
. "onsequently certain fatty acids such as arachidonic acid
cannot be made @from scratch@ (de no&o) in humans and other mammals. 1o$e&er
linoleic acid $hich plants ma*e can be con&erted to arachidonic acid. Arachidonate and
eicosapentaenoate are +.%carbon prostanoic acids that are the starting point of the
synthesis of prostaglandins thromboxanes and leu*otrienes.
-5.
/hich one of the follo$ing steps results in the formation of a phospholipid0
A. )tep A
!. )tep !
". )tep "
D. )tep D
#. )tep #
The answer is: C
"eramide is the basic unit composing all sphingolipids $hich include sphingomyelin and
gangliosides. )phingomyelin $hich usually contains phosphocholine as a polar head
group is the only phospholipid that does not ha&e a glycerol bac*bone. ;n contrast
gangliosides ha&e complex oligosaccharide head groups.
-,.
/hich of the follo$ing lipoproteins $ould contribute to a measurement of plasma
cholesterol in a normal person follo$ing a 1+%h fast0
A. Dery%lo$%density lipoproteins
!. 1igh%density lipoproteins
". "hylomicrons
D. "hylomicron remnants
#. Adipocyte lipid droplets
The answer is: B
;n the postabsorptional (postprandial) state plasma contains all the lipoproteinsG
chylomicrons deri&ed from dietary lipids pac*aged in the intestinal epithelial cells and
their remnants8 &ery%lo$%density lipoproteins (D>D>s) $hich contain endogenous lipids
and cholesterol pac*aged in the li&er8 lo$%density lipoproteins (>D>s) $hich are end
products of delipidation of D>D>s8 and high%density lipoprotins (1D>s) $hich are
synthesi6ed in the li&er. 1D>s are in part catalytic since transfer of their ";;
apolipoprotein to D>D>s or chylomicrons acti&ates lipoprotein lipase. ;n normal patients
only >D>s and 1D>s remain in plasma follo$ing a 1+%h fast since both chylomicrons
and D>D>s ha&e been delipidated. (ost of the cholesterol measured in blood plasma at
this time is present in the cholesterol%rich >D>s. 1o$e&er 1D> cholesterol also
contributes to the measurement.
-:.
A 1:%year%old boy has a long history of school problems and is labeled as hyperacti&e.
1is tissues are puffy gi&ing his face a @coarse@ appearance. 1is ;S tests ha&e declined
recently and are no$ mar*edly belo$ normal. >aboratory studies demonstrate normal
amounts of sphingolipids in fibroblast cultures $ith increased amounts of
glycosaminoglycans in urine. /hich of the follo$ing en6yme deficiencies might explain
the boyAs phenotype0
A. 1exosaminidase A B. 'lucocerebrosidase C. %>%iduronidase D. %
galactocerebrosidase E. %gangliosidase A
The answer is: C
The t$o major groups of lysosomal storage disease are sphingolipidoses and
mucopolysaccharidoses. An absence of %>%iduronidase as in 1urlerAs syndrome
(+:+E..) and )cheieAs syndrome (+:+E..) leads to accumulations of dermatan sulfate
and heparan sulfate. )cheieAs syndrome is less se&ere $ith corneal clouding joint
degeneration and increased heart disease. 1urlerAs syndrome has the same symptoms
plus mental and physical retardation leading to early death. The later onset in this child is
compatible $ith a diagnosis of )cheieAs syndrome. Note that 1urlerAs and )cheieAs
syndromes result from mutations at the same locus4hence their identical (cQusic*
numbers. The reasons for the differences in disease se&erity are un*no$n. All of the other
en6yme deficiencies listed lead to the lac* of proper brea*do$n of sphingolipids and
their accumulation as gangliosides glucocerebrosides and sphingomyelins. )ymptoms of
lipidoses may include organ enlargement mental retardation and early death.
-9.
>eu*ocyte samples isolated from the blood of a ne$born infant are homogeni6ed and
incubated $ith ganglioside '(+. Approximately ,BN of the expected normal amount of
*-acetylgalactosamine is liberated during the incubation period. These results indicate
that the infant
A. ;s a hetero6ygote (carrier) for Tay%)achs disease
!. ;s homo6ygous for Tay%)achs disease
". 1as Tay%)achs syndrome
D. /ill most li*ely ha&e mental deficiency
#. 1as relati&ely normal %N%acetylhexosaminidase acti&ity
The answer is: A
'angliosides are continually synthesi6ed and bro*en do$n. The specific hydrolases that
degrade gangliosides by sequentially remo&ing terminal sugars are found in lysosomes.
;n the lipid storage disease *no$n as Tay%)achs disease (+B+E..) ganglioside '(+
accumulates because of a deficiency of -*-acetylhexosaminidase a lysosomal en6yme
that remo&es the terminal *-acetylgalactosamine residue. 1omo6ygotes produce &irtually
no functional en6yme and suffer $ea*ness retardation and blindness. Death usually
occurs before infants are 5 years old. "arriers (hetero6ygotes) of the autosomal recessi&e
disease produce approximately :.N of the normal le&els of en6yme but sho$ no ill
effects. ;n high%ris* populations such as Ash*ena6i Pe$s screening for carrier status may
be performed.
-B.
;n humans the formation of the fatty acid "%1E%
-

1+
can be deri&ed from $hich of the
follo$ing0 A. "%1E cis%
-
B. "%1E cis-
9
C. "%1E D. "%19 cis-
9

-
E. "%
1E cis-
-

1+
The answer is: E
;n mammals a &ariety of fatty acids are considered essential and cannot be synthesi6ed.
These include linoleate ("%1E cis-
-

1+
) and linolenate ("%1E cis-
-

1+

1:
). #ither
these fatty acids or fatty acids for $hich they are precursors must be supplied in the diet
as starting points for synthesis of a &ariety of other unsaturated fatty acids that lead to the
synthesis of prostaglandins thromboxanes and leu*otrienes. =or example arachidonate
a +.%carbon fatty acid $ith four double bonds is deri&ed from linolenate. Arachidonate
gi&es rise to some prostaglandins thromboxanes and leu*otrienes. )ome fatty acids must
be obtained in the diet because of the limitations go&erning en6ymes of fatty acid
synthesis in humans8 that is double bonds cannot be introduced beyond the -J1. bond
position of carbons in the fatty acid chain and subsequent double bonds after the first
must be separated by t$o single bonds. Thus linolenate and linoleate cannot be
synthesi6ed in humans.
-E.
A ,:%year%old man has a mild heart attac* and is placed on diet and me&astatin therapy.
/hich of the follo$ing $ill be a result of this therapy0
A. >o$ blood glucose
!. >o$ blood >D>s
". 1igh blood cholesterol
D. 1igh blood glucose
#. >o$ oxidation of fatty acids
=. Qetosis
'. >ipolysis
The answer is: B
(e&astatin an analogue of me&alonic acid acts as a feedbac* inhibitor of 5A%hydroxy%5A%
methylglutaryl "oA (1('%"oA) reductase the regulated en6yme of cholesterol
synthesis. #ffecti&e treatment $ith me&astatin along $ith a lo$%fat diet decreases le&els
of blood cholesterol. The lo$ering of cholesterol also lo$ers the amounts of the
lipoprotein that transports cholesterol to the peripheral tissues lo$%density lipoprotein
(>D>). )ince lipids li*e cholesterol and triglycerides are insoluble in $ater they must be
associated $ith lipoproteins for transport and sal&age bet$een their major site of
synthesis (li&er) and the peripheral tissues. Those lipoproteins associated $ith more
insoluble lipids thus ha&e lo$er density during centrifugation a technique that separates
the lo$est%density chylomicrons from &ery%lo$%density lipoproteins (D>D>s $ith pre%
lipoproteins) lo$%density lipoproteins (>D>s $ith %lipoproteins) intermediate%density
lipoproteins (;D>s) and high%density lipoproteins (1D>s $ith %lipoproteins). #ach type
of lipoprotein has typical apolipoproteins such as the apo !1.. and apo !,E (translated
from the same messenger 2NA) in >D>. >D> is in&ol&ed in transporting cholesterol
from the li&er to peripheral tissues $hile 1D> is a sca&enger of cholesterol. The ratio of
1D> to >D> is thus a predictor of cholesterol deposition in blood &essels the cause of
myocardial infarctions (heart attac*s). The higher the 1D>M>D> ratio the lo$er the rate
of heart attac*s.
--.
A control and t$o patients $ith hyperlipidemia are studied after an o&ernight fast. Their
plasma lipoprotein electrophoresis patterns are sho$n belo$ the control being in the
middle lane. 7ne of the patients has a pattern typical of type ; lipoprotein lipase
deficiency and the other of type ;;a familial hypercholesterolemia. /hich of the bands
obser&ed in the electrophoretic gel patterns represents a lipoprotein fraction that is
abnormally abundant after fasting and that is most enriched in triacylglycerides0
A. !and A
!. !and !
". !and "
D. !and D
#. !and #
=. !and =
'. !and '
1. !and 1
;. !and ;
The answer is: A
3atients $ith type ; lipoprotein lipase deficiency are not able to rapidly delipidate
chylomicrons $hich carry dietary triacylglycerides or D>D>s $hich carry lipids
pac*aged by the li&er. The electrophoretic pattern (left lane in the figure) after fasting is
thus similar to that of a normal patient after a meal4the chylomicrons (band A) >D>s
$ith %lipoproteins (band !) D>D>s $ith pre% %lipoproteins (band ") and 1D>s $ith %
lipoproteins (band D) are all present because chylomicrons and D>D>s are not degraded
normally. 1epatic lipase ordinarily released by the li&er to deal $ith chylomicron
remnant and 1D> metabolism must slo$ly deal $ith the lipemia (lipoprotein buildup in
blood) caused by the lac* of lipoprotein lipase. ;n addition to the lipemia steatorrhea
(fatty stools) and stomach cramps are symptoms of lipoprotein lipase deficiency. A $ell%
controlled lo$%fat diet is part of the therapy. ;n contrast the electrophoretic pattern of
patients $ith type ;;a familial hypercholesterolemia <1,5E-. (right lane)? can appear at
first glance to be normal since only the >D> and 1D> bands are expected follo$ing an
o&ernight fast. 1o$e&er closer inspection re&eals an abnormally high accumulation of
>D>s (band ') $hich cause the hypercholesterolemia. )ince >D>s are a final stage in
the catabolism of D>D>s the high load of cholesterol being transported from the li&er
may cause some D>D>s to be present after fasting (band 1) along $ith 1D> sca&enger
lipoproteins (band ;). ;n patients hetero6ygous for the disease approximately one%half the
normal amount of !1.. >D> receptors are present. ;n homo6ygous patients no !1..
>D> receptors are present and most (UB.N) of the >D> must be cleared by the li&er. ;n
such patients blood cholesterol le&els are extraordinarily high and lead to profound early
atherosclerosis and death.
1...
A ,%year%old girl presents in the clinic $ith megaloblastic anemia and failure to thri&e.
!lood chemistries re&eal orotic aciduria. #n6yme measurements of $hite blood cells
re&eal a deficiency of the pyrimidine biosynthesis en6yme orotate
phosphoribosyltransferase and abnormally high acti&ity of the en6yme aspartate
transcarbamoylase. /hich one of the follo$ing treatments $ill re&erse all symptoms if
carried out chronically0
A. !lood transfusion
!. /hite blood cell transfusion
". Dietary supplements of phosphoribosylpyrophosphate (3233)
D. 7ral thymidine
#. 7ral uridine
The answer is: E
7rotic aciduria is the buildup of orotic acid due to a deficiency in one or both of the
en6ymes that con&ert it to C(3. #ither orotate phosphoribosyltransferase and orotidylate
decarboxylase are both defecti&e or the decarboxylase alone is defecti&e. C(3 is the
precursor of CT3 "T3 and T(3. All of these end products normally act in some $ay to
feedbac*%inhibit the initial reactions of pyrimidine synthesis. )pecifically the lac* of
"T3 inhibition allo$s aspartate transcarbamoylase to remain highly acti&e and ultimately
results in a buildup of orotic acid and the resultant orotic aciduria. The lac* of "T3 T(3
and CT3 leads to a decreased erythrocyte formation and megaloblastic anemia. Cridine
treatment is effecti&e because uridine can easily be con&erted to C(3 by omnipresent
tissue *inases thus allo$ing CT3 "T3 and T(3 to be synthesi6ed and feedbac*%inhibit
further orotic acid production.
1.1.
/hich of the follo$ing $ould rule out hyperuricemia in a patient0
A. >esch%Nyhan syndrome
!. 'out
". Oanthine oxidase hyperacti&ity
D. "arbamoyl phosphate synthase deficiency
#. 3urine o&erproduction secondary to Don 'ier*eAs disease
The answer is: D
"arbamoyl phosphate ("A3) synthase ; is found in mitochondrial matrix and is the first
step in urea synthesis condensing "7+ and N1,
I
. 1yperammonemia occurs $hen "A3 is
deficient. "A3 synthase ;; forms "A3 as the first step in pyrimidine synthesis. ;ts
complete deficiency $ould probably be a lethal mutation. /hen its acti&ity is decreased
purine catabolism to uric acid is decreased decreasing the possibility of hyperuricemia.
;n contrast gout >esch%Nyhan syndrome high xanthine oxidase acti&ity and &on
'ier*eAs disease <glycogen storage disease type ;a (+5++..)? all lead to increased urate
production and excretion.
1.+.
/hich one of the follo$ing contributes nitrogen atoms to both purine and pyrimidine
rings0
A. Aspartate
!. "arbamoyl phosphate
". "arbon dioxide
D. 'lutamine
#. Tetrahydrofolate
The answer is: A
During purine ring biosynthesis the amino acid glycine is completely incorporated to
pro&ide ", ": and NB. 'lutamine contributes N5 and N- aspartate pro&ides N1 and
deri&ati&es of tetrahydrofolate furnish "+ and "E. "arbon dioxide is the source of "9. ;n
pyrimidine ring synthesis "+ and N5 are deri&ed from carbamoyl phosphate $hile N1
", ": and "9 come from aspartate.
a % "ontrol of the rate of de no&o purine synthesis. )olid lines represent metabolite flo$
and dashed lines feedbac* inhibition ( ) by end products of the path$ay.
(Reproduced, with permission, from Murray RK, ranner !K, Mayes "#, Rodwell V$%
1arperAs !iochemistry &'(e) *ew Yor+, Mcraw-,ill, &---% 34.)1
b % 2egulation of the con&ersion of ;(3 to adenosine and guanosine nucleotides. )olid
lines represent metabolite flo$ and dashed lines represent positi&e ( ) or negati&e ( )
feedbac* inhibition.
1.5.
=eedbac* inhibition of pyrimidine nucleotide synthesis can occur by $hich of the
follo$ing means
A. ;ncreased acti&ity of carbamoyl phosphate synthetase
!. ;ncreased acti&ity of aspartate transcarbamoylase
". "T3 allosteric effects
D. C(3 competiti&e inhibition
#. TT3 allosteric effects
The answer is: C
The steps of pyrimidine nucleotide biosynthesis are summari6ed in the figure belo$. The
first step in pyrimidine synthesis is the formation of carbamoyl phosphate. The en6yme
cataly6ing this step carbamoyl phosphate synthetase (1) is feedbac*%inhibited by C(3
through allosteric effects on en6yme structure (not by competiti&e inhibition $ith its
substrates). The en6yme of the second step aspartate transcarbamoylase is composed of
catalytic and regulatory subunits. The regulatory subunit binds "T3 or AT3. TT3 has no
role in the feedbac* inhibition of pyrimidine synthesis. Decreased rather than increased
acti&ity of en6ymes 1 and + $ould be produced by allosteric feedbac* inhibition.
1.,.
3urine nucleotide biosynthesis can be inhibited by $hich of the follo$ing0
A. 'uanosine triphosphate ('T3)
!. Cridine monophosphate (C(3)
". Adenosine monophosphate (A(3)
D. Adenosine triphosphate (AT3)
#. ;nosine diphosphate (;D3)
The answer is: C
)e&eral control sites exist in the path of purine synthesis $here feedbac* inhibition
occurs. A(3 '(3 or ;(3 may inhibit the first step of the path$ay $hich is the
synthesis of :%phosphoribosyl%1%pyrophosphate (3233). 3233 synthetase is specifically
inhibited. All three nucleotides can inhibit glutamine 3233 aminotransferase $hich
cataly6es the second step of the path$ay. A(3 bloc*s the con&ersion of ;(3 to
adenylo@succinate. '(3 inhibits the formation of xanthylate from ;(3. Thus bloc*age
rather than enhancement of ;(3 metabolism to A(3 and '(3 effecti&ely inhibits purine
biosynthesis.
1.:.
/hich of the follo$ing compounds is a required substrate for purine biosynthesis0
A. :%methyl thymidine
!. Ara "
". 2ibose phosphate
D. :%phosphoribosylpyrophosphate (3233)
#. :%fluorouracil
The answer is: C
:A%phosphoribosyl%1%pyrophosphate (3233) donates the ribose phosphate unit of
nucleotides and is absolutely required for the beginning of the synthesis of purines. ;n
fact the en6ymes regulating the synthesis of 3233 and the subsequent synthesis of
phosphoribosylamine from 3233 are all end productJinhibited by inosine
monophosphate (;(3) adenosine monophosphate (A(3) and guanosine
monophosphate ('(3) the products of this reaction path$ay. Allopurinol an analogue
of hypoxanthine is a drug used to correct gout. ;t accomplishes this by inhibiting the
production of urate from hypoxanthine and in doing so undergoes suicide inhibition of
xanthine oxidase. :%fluorouracil is an analogue of thymidine that inhibits thymidylate
synthetase and is used in cancer chemotherapy. "ytosine arabinoside (Ara ") an inhibitor
of 2NA synthesis also ta*es ad&antage of rapid nucleic acid synthesis in cancer cells and
is used in chemotherapy.
1.9.
/hich of the follo$ing compounds is an analogue of hypoxanthine0
A. Ara "
!. Allopurinol
". 2ibose phosphate
D. :%phosphoribosylpyrophosphate (3233)
#. :%=C
The answer is: B
The degradation of purines to urate can lead to gout $hen an ele&ated le&el of urate is
present in serum causing the precipitation of sodium urate crystals in joints. The
excessi&e production of urate in many patients seems to be connected to a partial
deficiency of hypoxanthine%guanine phosphoribosyl transferase (1'32T). Allopurinol
an analogue of hypoxanthine is a drug used to correct gout. ;t accomplishes this by
inhibiting the production of urate from hypoxanthine and in doing so undergoes suicide
inhibition of xanthine oxidase. 2ibose phosphate and 3233 are required for purine
synthesis. :%fluorouracil (:%=C) and cytosine arabinoside (Ara ") are cancer
chemotherapy agents the former being an analogue of thymine that inhibits thymidylate
synthetase and the latter an inhibitor of 2NA synthesis.
1.B.
A pentose $ith a :A%phosphate group a +A%hydroxyl group and a 1A%pyrimidine group
describes $hich of the follo$ing structures0
A. "ytosine
!. 'uanosine
". Thymidine
D. Thymidylate
#. "ytidylate
The answer is: E
7nly cytidylate ("(3) has a :A%phosphate group a ribose (pentose) group $ith a +A%
hydroxyl and a pyrimidine base. T$o purines <adenine (A) and guanine (')? and t$o
pyrimidines <cytosine (") and thymine (T)? occur in DNA. ;n 2NA uracil (C) replaces
thymine. 1ypoxanthine and inosine are biosynthetic precursors to the bases in nucleic
acids. !ases form nucleosides through bonding of a nitrogen $ith the "1 carbon (1A%
carbon) of a pentose sugar. Adenosine guanosine and cytidine can form ribonucleotides
$ith ribose (+A% and 5A%hydroxyl groups) or deoxyribonucleotides $ith deoxyribose (5A%
hydroxyl group only). Cridine occurs only as the ribonucleoside thymidine as the
deoxyribonucleotide (actually as thymidylate deoxyribonucleotide synthesi6ed from
uridylate by thymidylate sythetase). 2ibonucleotides (adenylate guanidylate cytidylate
uridylate) ha&e a phosphate ester on the :A%hydroxyl of ribose (note that the ri5o- prefix is
usually omitted). Deoxyribonucleotides ha&e a phosphate ester on the :A%hydroxyl of
deoxyribose (deoxyadenylate deoxyguanidylate deoxycytidylate and thymidylate).
Thymidylate does not need the deoxy- prefix since it only forms as a
deoxyribonucleotide. The phosphate ester may be formed of one t$o or three phosphate
groups (e.g. A(3 AD3 or AT3).
1.E.
/hich of the acti&ated groups or units is most closely associated $ith uridine
diphosphate (CD3)0
A. #lectrons
!. 3hosphoryl
". Acyl
D. Aldehyde
#. 'lucose
The answer is: E
The acti&ated form of glucose utili6ed for the synthesis of glycogen and galactose is
CD3%glucose $hich is formed from the reaction of glucose%1%phosphate and CT3. The
&itamin lipoic acid is co&alently bound to the %amino group of a lysine residue of the
en6yme dihydrolipoyl transacetylase. The amide%lin*ed lipolysine residue is *no$n as
lipoamide and is an acti&ated carrier of acyl groups deri&ed from the hydroxyethyl
deri&ati&e of thiamine pyrophosphate. ;n this manner lipoamide functions as one of the
coen6ymes in oxidati&e decarboxylation reactions. ;n reducti&e synthesis such as fatty
acid synthesis NAD31 is the major electron donor. This may be contrasted to NAD1
$hich is utili6ed for the generation of AT3 &ia electron transport.
1.-.
1umans most easily tolerate a lac* of $hich of the follo$ing nutrients0
A. 3rotein
!. ;odine
". "arbohydrate
D. >ipid
#. "alcium
The answer is: C
"ertain amino acids and lipids are dietary necessities because humans cannot synthesi6e
them. The energy usually obtained from carbohydrates can be obtained from lipids and
the con&ersion of some amino acids to intermediates of the citric acid cycle. These
alternati&e substrates can thus pro&ide fuel for oxidation and energy plus reducing
equi&alents for biosynthesis. ;odine is important for thyroid hormone synthesis $hile
calcium is essential for muscle contraction and bone metabolism.
11..
/hich of the follo$ing &itamins is the precursor of "oA0
A. 2ibofla&in
!. 3antothenate
". Thiamine
D. "obamide
#. 3yridoxamine
The answer is: B
3antothenate is the precursor of "oA $hich participates in numerous reactions
throughout the metabolic scheme. "oA is a central molecule of metabolism in&ol&ed in
acetylation reactions. Thus a deficiency of pantothenic acid $ould ha&e se&ere
consequences. There is no documented deficiency state for pantothenate ho$e&er since
this &itamin is common in foodstuffs.
111.
/hich one of the follo$ing transfers acyl groups0
A. Thiamine pyrophosphate
!. >ipoamide
". AT3
D. NAD1
#. =AD1
The answer is: B
>ipoamide li*e "oA transfers acetyl groups but it is a catalytic cofactor in an en6yme
complex rather than a stoichiometric cofactor li*e "oA. A reacti&e disulfide of lipoamide
lin*s the acetyl group to be transferred. >ipoamide becomes acetyllipoamide and then
dihydrolipoamide as it first accepts and then transfers an acyl group. This reaction and the
regeneration of lipoamide are cataly6ed by different parts of the dehydrogenase en6yme
complexes of pyru&ate dehydrogenase and isocitrate dehydrogenase. AT3 transfers
phosphoryl groups and thiamine pyrophosphate transfers aldehyde groups. NAD1 and
=AD1 transfer protons.
11+.
/hich one of the follo$ing cofactors must be utili6ed during the con&ersion of acetyl
"oA to malonyl "oA
A. Thiamine pyrophosphate
!. Acyl carrier protein (A"3)
". NAD1
D. !iotin
#. =AD
The answer is: D
The *ey en6ymatic step of fatty acid synthesis is the carboxylation of acetyl "oA to form
malonyl "oA. The carboxyl of biotin is co&alently attached to an %amino acid group of a
lysine residue of acetyl "oA carboxylase. The reaction occurs in t$o stages. ;n the first
step a carboxybiotin is formedG
1"75
J
I biotin%en6yme I AT3 "7+%biotin%en6yme I AD3 I 3i
;n the second step the "7+ is transferred to acetyl "oA to produce malonyl "oAG
"7+%biotin%en6yme I acetyl "oA malonyl "oA I biotin%en6yme
None of the other cofactors listed are in&ol&ed in this reaction.
115.
"oen6ymes deri&ed from the &itamin sho$n belo$ are required by en6ymes in&ol&ed in
the synthesis of $hich of the follo$ing0
A. AT3
!. CT3
". "T3
D. NAD1
#. NAD31
The answer is: A
The structure sho$n in the question is the &itamin folic acid. Tetrahydrofolic acid the
acti&e cofactor deri&ed from folic acid is required in t$o steps of purine synthesis and
thus required in the de no&o synthesis of AT3 and 'T3. "T3 and TT3 are pyrimidine base
deri&ati&es and although de no&o synthesis of the pyrimidine ring does not require
tetrahydrofolate the formation of thymine from uracil does. NAD1 and NAD31 require
niacin for their synthesis.
11,.
/hich of the follo$ing is a coen6yme0
A. 'lucose%9%phosphate
!. 'lucose%1%phosphate
". "alcium ion
D. >ipoic acid
#. CD3%glucose
The answer is: D
A coen6yme is a nonprotein organic molecule that binds to an en6yme to aid in its
catalytic function. Csually it is in&ol&ed in the transfer of a specific functional group. A
coen6yme usually binds loosely and can be separated from the en6yme. /hen a
coen6yme binds tightly to an en6yme it is spo*en of as a prosthetic group of the en6yme.
"oen6ymes can be &ie$ed as a second substrate for the en6yme often undergoing
chemical changes that counterbalance those of the substrate. >ipoic acid is a short%chain
fatty acid $ith t$o sulfhydryl groups. >ipoic acid is a coen6yme in&ol&ed in pyru&ate
dehydrogenase and %*etoglutarate dehydrogenase reactions ta*ing part in the reactions.
#ach of these reactions also uses thiamine pyrophosphate "oA =AD
I
and NAD
I
.
'lucose%1%phosphate glucose%9%phosphate or CD3%glucose function as primary
substrates of en6ymatic reactions andMor induce structural changes in the en6yme
(allosteric regulators) to influence en6yme acti&ity. "alcium and other metal ions do not
undergo chemical changes during the en6yme reaction and are classified as cofactors.
/hile many coen6ymes are deri&ed by modification of &itamins such as "oA from
pantothenic acid or =AD from ribofla&in some coen6ymes do not contain &itamins. =or
example ubiquinone (oxidi6ed) or ubiquinol (reduced) is in fact coen6yme S $hich is
in&ol&ed in transferring hydrogen ion atoms and electrons in the oxidati&e
phosphorylation system. )ince humans can synthesi6e ubiquinones and lipoic acid these
substances are not considered &itamins.
11:.
A term infant is born at home and does $ell $ith breast%feeding. T$o days later the
mother calls frantically because the baby is bleeding from the umbilical cord and nostrils.
/hich of the follo$ing is the most li*ely cause of the bleeding0
A. Deficiency of &itamin " due to a citrus%poor diet during pregnancy
!. 1yper&itaminosis A due to ingestion of beef li&er during pregnancy
". Deficiency of &itamin Q because infant intestines are sterile
D. Deficiency of &itamin Q because of disseminated intra&ascular coagulation
(disseminated clotting due to infantile sepsis)
#. Deficiency of &itamin # due to maternal malabsorption during pregnancy
The answer is: C
1emorrhagic disease of the ne$born is caused by poor transfer of maternal &itamin Q
through the placenta and by lac* of intestinal bacteria in the infant for synthesis of
&itamin Q. The intestine is sterile at birth and becomes coloni6ed o&er the first fe$
$ee*s. !ecause of these factors &itamin Q is routinely administered to ne$borns.
Deficiencies of the fat%soluble &itamins A # D and Q can occur $ith intestinal
malabsorption but a&id fetal upta*e during pregnancy usually pre&ents infantile
symptoms. 1yper&itaminosis A can cause li&er toxicity but not bleeding and deficiencies
of # (neonatal anemia) or " (extremely rare in neonates) ha&e other symptoms besides
bleeding.
119.
/hich of the follo$ing conditions most rapidly produces a functional deficiency of
&itamin Q0
A. "oumadin therapy to pre&ent thrombosis in patients prone to clot formation
!. !road%spectrum antibiotic therapy
". >ac* of red meat in the diet
D. >ac* of citrus fruits in the diet
#. 3remature birth
The answer is: A
Ditamin Q is essential for the posttranscriptional modification of prothrombin by %
carboxylation of glutamate residues. A functional deficiency exists in patients treated
$ith analogues of &itamin Q such as the coumadin deri&ati&es. The analogues act as
anticoagulants by competing $ith &itamin Q and pre&enting the production of functional
prothrombin. !y administration of &itamin Q hemorrhage can be pre&ented in such
patients. Ditamin Q is normally obtained from green leafy &egetables in the diet (not
from citrus fruits or red meat). ;ntestinal bacteria also synthesi6e the &itamin but e&en
broad%spectrum antibiotic therapy does not completely sterili6e the intestine. A deficiency
of &itamin Q can cause hemorrhage disease in ne$born infants since their intestines do
not ha&e the bacteria that produce &itamin Q and since &itamin Q does not cross the
placenta. The neonatal deficiency occurs in term or premature infants.
11B.
A 5%month%old boy presents $ith poor feeding and gro$th lo$ muscle tone (hypotonia)
ele&ation of blood lactic acid (lactic acidemia) and mild acidosis (blood p1 B.5 to B.5:).
The ratio of pyru&ate to lactate in serum is ele&ated and there is decreased con&ersion of
pyru&ate to acetyl coen6yme A in fibroblasts. /hich of the follo$ing compounds might
be considered for therapy0
A. 3yridoxine
!. Thiamine
". =ree fatty acids
D. !iotin
#. Ascorbic acid
The answer is: B
An ele&ation of pyru&ate and a deficiency of acetyl "oA suggest a deficiency of pyru&ate
dehydrogenase (3D1). This multisubunit en6yme assembly contains pyru&ate
dehydrogenase dihydrolipoyl transacetylase dihydrolipoyl dehydrogenase and t$o
en6ymes in&ol&ed in regulation of the o&erall en6ymatic acti&ity of the complex. 3D1
requires thiamine pyrophosphate as a coen6yme dihydrolipoyl transacetylase requires
lipoic acid and "oA and dihydrolipoyl dehydrogenase has an =AD prosthetic group that
is reoxidi6ed by NAD
I
. !iotin pyridoxine and ascorbic acid are not coen6ymes for
3D1. An AT3%dependent protein *inase can phosphorylate 3D1 to decrease acti&ity and
a phosphatase can acti&ate 3D1. ;ncreases of AT3 acetyl "oA or NAD1 (increased
energy charge) and of fatty acid oxidation increase phosphorylation of 3D1 and decrease
its acti&ity. 3D1 is less acti&e during star&ation increasing pyru&ate decreasing
glycolysis and sparing carbohydrates. =ree fatty acids decrease 3D1 acti&ity and $ould
not be appropriate therapy for 3D1 deficiency. 3D1 deficiency (+,9-.. 51+1B.)
exhibits genetic heterogeneity as $ould be expected from its multiple subunits $ith
autosomal and O%lin*ed recessi&e forms. The infant also could be classified as ha&ing
>eighAs disease (+991:.) a heterogenous group of disorders $ith hypotonia and lactic
acidemia that can include 3D1 deficiency.
11E.
A homeless person is brought into the emergency room $ith psychotic imagery and
alcohol on his breath. /hich of the follo$ing compounds is most important to
administer0
A. 'lucose
!. Niacin
". Nicotinic acid
D. Thiamine
#. 2ibofla&in
The answer is: D
"hronic alcoholics are at ris* for thiamine deficiency $hich is thought to play a role in
the incoordination (ataxia) and psychosis that can become chronic (/ernic*e%Qorsa*off
syndrome). The thiamine deficiency produces relati&e deficiency of the pyru&ate
dehydrogenase complex. The administration of glucose $ithout chec*ing glucose le&els
can therefore be dangerous since excess glucose is con&erted to pyru&ate by glycolysis.
The lo$ rate of pyru&ate dehydrogenase con&ersion of pyru&ate to coen6yme A (and
entry into the citric acid cycle) causes pyru&ate to be con&erted to lactate (through lactate
dehydrogenase). >actic acidosis can be fatal. "hronic alcoholics can be deficient in the
other &itamins mentioned but thiamine is most li*ely to help the neurologic symptoms.
11-.
/hich of the follo$ing &itamins becomes a major electron acceptor aiding in the
oxidation of numerous substrates0
A. Ditamin !9
!. Niacin
". 2ibofla&in
D. Thiamine
#. Ditamin !
The answer is: B
Nicotinamide adenine dinucleotide (NAD
I
) is the functional coen6yme deri&ati&e of
niacin. ;t is the major electron acceptor in the oxidation of molecules generating NAD1
$hich is the major electron donor for reduction reactions. Thiamine (also *no$n as
&itamin !1) occurs functionally as thiamine pyrophosphate and is a coen6yme for
en6ymes such as pyru&ate dehydrogenase. 2ibofla&in (&itamin !+) functions in the
coen6yme forms of fla&in mononucleotide (=(N) or fla&in adenine dinucleotide (=AD).
/hen concentrated both ha&e a yello$ color due to the ribofla&in they contain. !oth
function as prosthetic groups of oxidation%reduction en6ymes or fla&oproteins.
=la&oproteins are acti&e in selected oxidation reactions and in electron transport but they
do not ha&e the ubiquitous role of NAD
I
.
1+..
/hich of the follo$ing &itamins can act $ithout phosphorylation0
A. 3yridoxine
!. >ipoamide
". Niacin
D. Thiamine
#. 2ibofla&in
The answer is: B
All the &itamins listed except lipoamide contain at least one phosphate in their cofactor
form. Thiamine (&itamin !1) is con&erted to thiamine pyrophosphate simply by the
addition of pyrophosphate. ;t is in&ol&ed in aldehyde group transfer. Niacin (nicotinic
acid) is esterified to adenine dinucleotide and its t$o phosphates to form nicotinamide
adenine dinucleotide. 3yridoxine (&itamin !9) is con&erted to either pyridoxal phosphate
or pyridoxamine phosphate before complexing $ith en6ymes. 2ibofla&in becomes fla&in
mononucleotide by obtaining one phosphate (ribofla&in :A%phosphate). ;f it complexes
$ith adenine dinucleotide &ia a pyrophosphate ester lin*age it becomes fla&in adenine
dinucleotide.
1+1.
A +%year%old child presents $ith chronic cough and bronchitis gro$th failure and
chronic diarrhea $ith light%colored foul%smelling stools. A deficiency of $hich of the
follo$ing &itamins should be considered0
A. Ditamin A
!. Ditamin "
". Ditamin !1
D. Ditamin !+
#. Ditamin !
The answer is: A
Ditamins A D # and Q are all fat%soluble. The physical characteristics of fat%soluble
&itamins deri&e from the hydrophobic nature of the aliphatic chains composing them. The
other &itamins listed are $ater%soluble efficiently administered orally and rapidly
absorbed from the intestine. =at%soluble &itamins must be administered intramuscularly
or as oral emulsions (mixtures of oil and $ater). ;n intestinal disorders such as chronic
diarrhea or malabsorption due to deficient digesti&e en6ymes fat%soluble &itamins are
poorly absorbed and can become deficient. )upplementation of fat%soluble &itamins is
thus routine in disorders li*e cystic fibrosis (+1-B..) a cause of respiratory and intestinal
disease that is the li*ely diagnosis in this child.
1++.
/hich of the follo$ing en6ymes requires a phosphorylated deri&ati&e of the &itamin
sho$n belo$0
A. 3yru&ate carboxylase
!. 3yru&ate dehydrogenase
". 3hosphoenolpyru&ate carboxy*inase
D. 'luco*inase
#. =ructo*inase
The answer is: B
The &itamin sho$n in the question is thiamine (&itamin !1) $hich requires
phosphorylation to thiamine pyrophosphate for acti&ity. Thiamine is required for the
reactions cataly6ed by pyru&ate dehydrogenase trans*etolases and %*etoglutarate
dehydrogenase. ;n all these reactions thiamine is in&ol&ed $ith oxidati&e
decarboxylation. The fi&e%member thia6ole ring of thiamine pyrophosphate forms a
carbanion (carbon bet$een the nitrogen and sulfur) that can react $ith "F7 groups
causing elimination of the carbonyl group ("771) as "7+. Qinases such as those in
glycolysis require AT3 as a cofactor $hile pyru&ate carboxylase and other carboxylases
require biotin to transfer an acti&ated carbonyl group.
1+5.
A deficiency of $hich of the follo$ing &itamins is associated $ith the occurrence of
neural tube defects (anencephaly and spina bifida)0
A. Ascorbic acid (&itamin ")
!. Thiamine (&itamin !1)
". 2ibofla&in (&itamin !+)
D. Niacin (&itamin !5
#. !iotin
=. 3antothenic acid
'. =olic acid
1. "obalamin (&itamin !1+)
The answer is: G
)pina bifida or myelomeningocele is a defect of the lo$er neural tube that produces an
exposed spinal cord in the thoracic or sacral regions. #xposure of the spinal cord usually
causes ner&e damage that results in paralysis of the lo$er limbs and urinary bladder.
Anencephaly is a defect of the anterior neural tube that results in lethal brain anomalies
and s*ull defects. =olic acid is necessary for the de&elopment of the neural tube in the
first fe$ $ee*s of embryonic life and the children of $omen $ith nutritional deficiencies
ha&e higher rates of neural tube defects. )ince neural tube closure occurs at a time $hen
many $omen are not a$are that they are pregnant it is essential that all $omen of
childbearing age ta*e a folic acid supplement of approximately .., mg per day. =ran*
folic acid deficiency can also cause megaloblastic anemia because of a decreased
synthesis of the purines and pyrimidines needed for cells to ma*e DNA and di&ide.
Deficiencies of thiamine in chronic alcoholics are related to /ernic*e%Qorsa*off
syndrome $hich is characteri6ed by loss of memory lac*adaisical beha&ior and a
continuous rhythmic mo&ement of the eyeballs. Thiamine dietary deficiency from excess
of polished rice can cause beriberi. Niacin deficiency leads to pellagra a disorder that
produces s*in rash (dermatitis) $eight loss and neurologic changes including depression
and dementia. 2ibofla&in deficiency leads to mouth ulcers (stomatitis) cheilosis (dry
scaly lips) scaly s*in (seborrhea) and photophobia. )ince biotin is $idely distributed in
foods and is synthesi6ed by intestinal bacteria biotin deficiency is rare. 1o$e&er the
heat%labile molecule a&idin found in ra$ egg $hites binds biotin tightly and bloc*s its
absorption causing dermatitis dehydration and lethargy. >actic acidosis results as a
buildup of lactate due to the lac* of functional pyru&ate carboxylase $hen biotin is
missing. Ditamin " deficiency leads to scur&y $hich causes bleeding gums and bone
disease.
Ditamin !1+ can be deficient due to a lac* of intrinsic factor $hich is a glycoprotein
secreted by gastric parietal cells. A lac* of intrinsic factor or a dietary deficiency of
cobalamin can cause pernicious anemia and neuropsychiatric symptoms. The only *no$n
treatment for intrinsic factor deficiency (&itamin !1+ deficiency) is intramuscular
injection of cyanocobalamin throughout the patientAs life.
1+,.
An African American infant presents $ith prominent forehead bo$ing of the limbs
broad and tender $rists s$ellings at the costochondral junctions of the ribs and
irritability. /hich of the follo$ing treatments is most appropriate0
A. >otions containing retinoic acid
!. Diet of baby food containing leafy &egetables
". Diet of baby food containing li&er and ground beef
D. (il* and sunlight exposure
#. 2emo&al of eggs from diet
The answer is: D
3eople $ith bo$ed legs and other bone malformations $ere quite common in the
northeastern Cnited )tates follo$ing the industrial re&olution. This $as caused by
childhood diets lac*ing foods $ith &itamin D and by minimal exposure to sunlight due to
the da$n%to%dus* $or*ing conditions of the textile mills. Ditamin D is essential for the
metabolism of calcium and phosphorus. )oft and malformed bones result from its
absence. >i&er fish oil and egg yol*s contain &itamin D and mil* is supplemented $ith
&itamin D by la$. ;n adults lac* of sunlight and a diet poor in &itamin D lead to
osteomalacia (soft bones). Dar*%s*inned peoples are more susceptible to &itamin D
deficiency.
!iotin deficiency can be caused by diets $ith excess egg $hite leading to dehydration
and acidosis from accumulation of carboxylic and lactic acids. 2etinoic acid is a &itamin
A deri&ati&e that can be helpful in treating acne but not &itamin D deficiency. >eafy
&egetables are a source of ! &itamins such as niacin and cobalamin.
1+:.
/hich of the follo$ing is noted in "ushingAs syndrome a disease of the adrenal cortex0
A. Decreased production of epinephrine
!. #xcessi&e production of epinephrine
". #xcessi&e production of &asopressin
D. #xcessi&e production of cortisol
#. Decreased production of cortisol
The answer is: D
A tumor of the adrenal cortex $ould be expected to affect the production of adrenal
steroid hormone. "ortisol and aldosterone are synthesi6ed in the cortex. ;n "ushingAs
syndrome hypersecretion of cortisol occurs. "ortisol is a glucocorticoid that has the
effect of encouraging metabolism of proteins lipids and carbohydrates. ;n some cases of
"ushingAs disease excessi&e production of cortisol is due to high le&els of A"T1
produced as a result of pituitary tumors. Diseases affecting the adrenal medulla might be
expected to disrupt or potentiate production of epinephrine in some $ay. #pinephrine
(adrenalin) is synthesi6ed in the medulla.
1+9.
;ncreased reabsorption of $ater from the *idney is the major consequence of $hich of the
follo$ing hormones0
A. "ortisol
!. ;nsulin
". Dasopressin
D. 'lucagon
#. Aldosterone
The answer is: C
Dasopressin $hich is also called antidiuretic hormone increases the permeability of the
collecting ducts and distal con&oluted tubules of the *idney and thus allo$s passage of
$ater. >i*e the mineralocorticoid aldosterone &asopressin results in an expansion of
blood &olume. 1o$e&er the mode of action of aldosterone is different8 it causes sodium
reabsorption not $ater reabsorption. )odium reabsorption indirectly leads to increased
plasma osmolality and thus $ater retention in the blood. "ortisol is a glucocorticoid that
potentiates catabolic metabolism chronically. #pinephrine stimulates catabolic
metabolism acutely. ;nsulin acutely fa&ors anabolic metabolism in large part by allo$ing
glucose and amino acid transport into cells.
1+B.
>ac* of glucocorticoids and mineralocorticoids might be a consequence of $hich of the
follo$ing defects in the adrenal cortex0
A. Androstenedione deficiency B. 1B %hydroxyprogesterone deficiency C.
#strone deficiency D. "%+1%hydroxylase deficiency E. Testosterone deficiency
The answer is: D
!oth cortisol and aldosterone contain "%+1%hydroxyl groups. !oth are also deri&ed from
progesterone in the adrenal cortex. ;n contrast the sex hormones are synthesi6ed in the
o&aries and testicular interstitial cells. ;n the synthesis of sex hormones progesterone is
con&erted to 1B %hydroxyprogesterone and then androstenedione $hich may either
become estrone or testosterone. Testosterone gi&es rise to estradiol in the o&aries. ;n the
corpus luteum progesterone is produced.
1+E.
A patient presents $ith a complaint of muscle $ea*ness follo$ing exercise. Neurological
examination re&eals that the muscles supplied by cranial ner&es are most affected. Rou
suspect myasthenia gra&is. Rour diagnosis is confirmed $hen lab tests indicate antibodies
in the patientAs blood against
A. Acetylcholinesterase
!. (uscle endplates
". "ranial ner&e synaptic membranes
D. "ranial ner&e presynaptic membranes
#. Acetylcholine receptors
The answer is: E
The major problem in myasthenia gra&is is a mar*ed reduction of acetylcholine receptors
on the motor endplate $here cranial ner&es form a neuromuscular junction $ith muscles.
;n these patients autoantibodies against the acetylcholine receptors effecti&ely reduce
receptor numbers. Normally acetylcholine molecules released by the ner&e terminal bind
to receptors on the muscle endplate resulting in a stimulation of contraction by
depolari6ing the muscle membrane. The condition is impro&ed $ith drugs that inhibit
acetylcholinesterase.
1+-.
/hich of the follo$ing hormones can cause hyperglycemia $ithout *no$n effects on
glycogen or gluconeogenesis0
A. Thyroxine
!. #pinephrine
". 'lucocorticoids
D. #pidermal gro$th factor
#. 'lucagon
The answer is: A
Thyroid hormones raise serum glucose le&els but the mechanism is un*no$n.
#pinephrine and glucagon lead to acute hyperglycemic effects by acti&ating li&er
phosphorylase and the release of glucose from glycogen through a cyclic A(3 cascade
effect. 'lucocorticoids stimulate the li&er to produce more gluconeogenic en6ymes and
promote protein brea*do$n to form amino acids. 'lucocorticoid (a steroid hormone) and
thyroxine exert chronic effects and act by binding to intracellular binding proteins
(receptors) that e&entually act as enhancers of transcription. #pidermal gro$th factor
receptor is similar to the insulin receptor in that it has a tyrosine *inase acti&ity that is
acti&ated by the binding of gro$th factor to the extracellular portion of the protein.
#pidermal gro$th factor does not cause hyperglycemia.
15..
=ollo$ing release of norepinephrine by sympathetic ner&es and epinephrine by the
adrenal medulla $hich of the follo$ing metabolic processes is decreased0
A. 'lycolysis
!. >ipolysis
". 'luconeogenesis
D. 'lycogenolysis
#. Qetogenesis
The answer is: A
The actions of epinephrine (adrenaline) and norepinephrine are catabolic8 that is these
catecholamines are antagonistic to the anabolic functions of insulin and li*e glucagon
are secreted in response to lo$ blood glucose or during @fight or flight@ stress. 'lycolysis
is an anabolic process that is decreased in the presence of ele&ated catecholamines.
Cnli*e glucagon $hich only acts on the li&er the catecholamines affect most tissues
including li&er and muscle. The catabolic processes increased by secretion of epinephrine
and norepinephrine include glycogenolysis gluconeogenesis lipolysis and *etogenesis.
Thus products that increase blood sugar or spare it such as *etone bodies and fatty acids
are increased.
151.
/hich step in the diagram belo$ is thought to be responsible for the effect of anti%
inflammatory steroids0
A. )tep A
!. )tep !
". )tep "
D. )tep D
#. )tep #
The answer is: A
The e&idence indicates that anti%inflammatory steroids inhibit phospholipase A+ $hich is
responsible for hydroly6ing arachidonate off of membrane phospholipids. "orticosteroids
and their manufactured deri&ati&es are thought to cause induction of the phospholipase
A+Jinhibitory protein lipocortin. ;n this manner production of all of the deri&ati&es of
arachidonic acid (lipoxins leu*otrienes thromboxanes and prostaglandins) is shut off. ;n
contrast nonsteroidal anti%inflammatory agents such as aspirin indomethacin and
ibuprofen act by inhibiting the cyclooxygenase component of prostaglandin synthase. The
synthase is responsible for the first step in the production of prostaglandins (step #) and
thromboxanes (step D) from arachidonic acid. The lipoxygenase path$ay leads to the
synthesis of lipoxins (step !) and leu*otrienes (step ").
15+.
/hich of the follo$ing effects of the steroid digitalis is obser&ed after treatment of
congesti&e heart failure0
A. Decrease in cytosolic sodium le&els
!. ;nhibition of NaIQI%AT3ase
". Decrease in the force of heart muscle contraction
D. )timulation of the plasma membrane ion pump
#. Decrease in cytosolic calcium
The answer is: B
Treatment of patients $ith congesti&e heart failure is often based on the use of
cardiotonic steroids such as digitalis. Digitalis is deri&ed from the foxglo&e plant and has
been used as an herbal treatment for heart problems since ancient times. Digitalis and
ouabain are cardiotonic steroids that inhibit the Na
I
Q
I
%AT3ase pump located in the
plasma membrane of cardiac muscle cells. They specifically inhibit the
dephosphorylation reaction of the AT3ase $hen the cardiotonic steroid is bound to the
extracellular face of the membrane. Due to inhibition of the pump higher le&els of
sodium are left inside the cell leading to a diminished sodium gradient. This results in a
slo$er exchange of calcium by the sodium%calcium exchanger. )ubsequently
intracellular le&els of calcium are maintained at a higher le&el and greatly enhance the
force of contraction of cardiac muscle.
155.
Aspirin inhibits $hich of the follo$ing en6ymes0
A. >ipoprotein lipase
!. >ipoxygenase
". "yclooxygenase
D. 3hospholipase D
#. 3hospholipase A+
The answer is: C
During the synthesis of prostaglandins a specific fatty acid is released from the +A
position of membrane phospholipids by the action of phospholipase A+. After its release
the fatty acid can enter either the lipoxygenase path$ay $hich produces acid $ith an
un*no$n biologic function or the prostaglandin cyclooxygenase (also called
prostaglandin synthetase) path$ay. ;n the formation of prostaglandins from fatty acids
cyclooxygenase cataly6es formation of a cyclopentane ring and the introduction of three
oxygen atoms. The type of prostaglandin produced depends on the starting fatty acid
$hich is al$ays a deri&ati&e of an essential fatty acid. #icosatrienoic acid yields series 1
prostaglandins eicosatetraenoic (arachidonic) acid yields series + prostaglandins and
eicosapentaenoic acid yields series 5 prostaglandins. Aspirin li*e indomethacin
decreases prostaglandin synthesis by inhibiting the oxygenase acti&ity of cyclooxygenase.
15,.
/hich of the follo$ing processes yields arachidonic (:E111,%eicosatetraenoic) acid in
mammals0
A. #longation of stearic acid
!. "hain elongation and one desaturation of linolenic (-1+1:%octadecatrienoic) acid
". "hain elongation and t$o desaturations of linoleic (-1+%octadecadienoic) acid
D. Desaturation of oleic acid
#. #longation of palmitic acid
The answer is: C
;n mammals arachidonic (:E111:%eicosatetraenoic) acid can only be synthesi6ed from
essential fatty acids deri&ed from the diet. >inoleic (-1+%octadecadienoic) acid produces
arachidonic acid follo$ing t$o desaturations and chain elongation. /hile linolenic
(-1+1:%octadecatrienoic) acid also is an essential fatty acid desaturation and elongation
produce E111,1B%eicosatetraenoic acid $hich is distinct from arachidonic acid. 7leic
palmitic and stearic acids are all nonessential fatty acids that cannot gi&e rise to
arachidonic acids in mammals.
15:.
A patient stung by a bee is rushed into the emergency room $ith a &ariety of symptoms
including increasing difficulty in breathing due to &asal and bronchial constriction. /hile
your subsequent treatment is to bloc* the effects of histamine and other acute%phase
reactants released by most cells you must also bloc* the slo$%reacting substance of
anaphylaxis ()2)%A) $hich is the most potent constrictor of the muscles en&eloping the
bronchial passages. /hat is )2)%A composed of0
A. Thromboxanes
!. ;nterleu*ins
". "omplement
D. >eu*otrienes
#. 3rostaglandins
The answer is: D
>eu*otrienes ", D, and #, together compose the slo$%reacting substance of anaphylaxis
()2)%A) $hich is thought to be the cause of asphyxiation in indi&iduals not treated
rapidly enough follo$ing an anaphylactic shoc*. )2)%A is up to 1... times more
effecti&e than histamines in causing bronchial muscle constriction. Anti%inflammatory
steroids are usually gi&en intra&enously to end chronic bronchoconstriction and
hypotension follo$ing a shoc*. The steroids bloc* phospholipase A+ action pre&enting
the synthesis of leu*otrienes from arachidonic acid. Acute treatment in&ol&es epinephrine
injected subcutaneously initially and then intra&enously. Antihistamines such as
diphenhydramine are administered intra&enously or intramuscularly.
159.
The central ring structure sho$n belo$ is found in $hich of the follo$ing compounds0
A. Adrenocorticotropin
!. Aldosterone
". 'eranyl phosphate
D. 3rostaglandin
#. Ditamin "
The answer is: B
)teroid hormones such as aldosterone are ultimate deri&ati&es of cholesterol. The
compound illustrated in the question is cholesterol one of a large group of steroids.
"holesterol $hich can be deri&ed from the diet as $ell as synthesi6ed de no&o is the
precursor of all steroids in&ol&ed in mammalian metabolism. These include the bile acids
the steroid hormones and &itamin D. "holesterol cannot be metaboli6ed to carbon
dioxide and $ater in humans. ;t must be excreted as a component of bile.
Adrenocorticotropin (A"T1) is a peptide hormone of the adenohypophysis that
influences the secretion of corticosteroid hormones. 3rostaglandins are eicosanoid
deri&ati&es that are also made up of isoprene units. 'eranyl phosphate is a +%isoprenoid
unit precursor in cholesterol synthesis.
15B.
/hich of the follo$ing compounds ser&es as a primary lin* bet$een the citric acid cycle
and the urea cycle0
A. (alate
!. )uccinate
". ;socitrate
D. "itrate
#. =umarate
The answer is: E
All the compounds listed are intermediates of the citric acid cycle. 1o$e&er only
fumarate is an intermediate of both the citric acid and urea cycles. ;t and arginine are
produced from argininosuccinate. 7nce produced by the urea cycle fumarate enters the
citric acid cycle and is con&erted to malate and then oxidi6ed to oxaloacetate. Depending
upon the organismAs needs oxaloacetate can either enter gluconeogenesis or react $ith
acetyl "oA to form citrate.
15E.
/hich one of the follo$ing can be con&erted to an intermediate of either the citric acid
cycle or the urea cycle0
A. Tyrosine
!. >ysine
". >eucine
D. Tryptophan
#. Aspartate
The answer is: E
Aspartate is a glucogenic amino acid that is also used to carry N1,
I
into the urea cycle.
Aspartate aminotransferase cataly6es the direct transamination of aspartate to
oxaloacetateG
aspartate I %*etoglutarate oxaloacetate I glutamate
7xaloacetate may either be utili6ed in the citric acid cycle or undergo gluconeogenesis.
Argininosuccinate synthetase cataly6es the condensation of citrulline and aspartate to
form argininosuccinateG
citrulline I asparate I AT3 argininosuccinate I A(3 I 33i
;n this manner one of the t$o nitrogens of urea is introduced into the urea cycle.

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