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MBBS FIRST PROFESSIONAL (PART II)

Biochemistry Theory
Max. Marks: 32.5 Time allowed: 2
hours

Q 1.
a. Describe any four inhibitors of oxidative phosphorylation along
with their specific mechanism and site of inhibition. (2)
b. Enumerate the steps of a polymerase chain reaction (PCR). Write
down the reactions of uric acid formation from hypoxanthine.
(1+1)
KEY
(a) Note: The following are the common inhibitors of electron transport chain,
however there may be other inhibitors. Only four of the following or other
compounds with such inhibitory action are required.
Name Mechanism of inhibition Site of Action
Rotenone e
-
transport inhibitor Complex I
Amytal e
-
transport inhibitor Complex I
Antimycin A e
-
transport inhibitor Complex III
Cyanide e
-
transport inhibitor Complex IV
Carbon Monoxide e
-
transport inhibitor Complex IV
Azide e
-
transport inhibitor Complex IV
2,4,-dinitrophenol Uncoupling agent
Transmembrane proton
carrier
Pentachlorophenol Uncoupling agent
transmembrane proton
carrier
Oligomycin Inhibits ATP synthase
OSCP fraction of ATP
synthase
(b)
Steps of a polymerase chain reaction:
1. Primer construction:
2. Denature the DNA
3. Annealing of primers to single-stranded DNA
4. Chain extension
Formation of uric acid from hypoxanthine
Hypoxanthine + O
2
+ H
2
O Xanthine + H
2
O
2
(catalyzed by xanthine oxidase)
Xanthine + H
2
O + O
2
Uric acid + H
2
O
2
(catalyzed by xanthine oxidase)
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Q 2.
a. Write down the irreversible reactions of glycolytic pathway. (1 )
b. What is the biochemical explanation for cataract formation,
retinopathy and peripheral neuropathy associated with
hyperglycemia in uncontrolled diabetes mellitus?. (2)

KEY
(a)
Glucose + ATP Glucose 6-phosphate + ADP (catalyzed by
glucokinase/hexokinase)
Fructose 6-phophate + ATP Fructose 1,6-bisphosphate + ADP (catalyzed by
phsophofructokinase 1)
Phosphoenol pyruvate + ADP Pyruvate + ATP (catalyzed by pyruvate kinase)

(b)
Insulin is not required for glucose entry into nerve cells, RBCs, etc. So large
amounts of glucose may enter these cells during times of hyperglycemia in
uncontrolled diabetes. Elevated intracellular glucose concentrations and an
adequate supply of NADPH cause aldose reductase to produce a significant
increase in the amount of sorbitol, which cannot pass efficiently through cell
membranes and, therefore, remains trapped inside the cell. Sorbitol
dehydrogenase becomes inadequate to metabolize the excess of sorbitol
accumulated in retina, lens, and nerve cells as a result of hyperglycemia, then it
causes strong osmotic effects and, therefore, cell swelling as a result of water
retention. Some of the pathologic alterations associated with diabetes can be
attributed, in part, to this phenomenon, including cataract formation,
retinopathy and peripheral neuropathy.

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Q 3.
a. Describe the Cori cycle and its importance. (2)
b. Name any four enzyme activities residing in fatty acid synthase
multienzyme complex. Name the enzyme defects in maple syrup
urine disease, albinism, phenylketonruia, and alkaptonuria.
(1+1)

KEY
(a)
In the Cori cycle, glucose is converted by
exercising muscle to lactate, which diffuses into
the blood. This lactate is taken up mainly by liver
and converted to glucose via gluconogenesis. The
glucose synthesized by liver via this metabolic
pathway enters blood circulation and plays an
important role in the maintenance of blood
glucose concentration.
(b)
Enzyme activities residing in fatty acid synthase
multienzyme complex (only four of the following
are required)
1. Ketoacyl synthase
2. Acetyl transacylase
3. Malonyl transacylase
4. Hydratase
5. Enoyl reductase
6. Ketoacyl reductase
7. Thioesterase

Disease Enzyme defect
Maple syrup urine
disease
Branched chain alpha-
ketoacid
dehydrogenase
Phenylketonuria Phenylalanine
hydroxylase
Albinism Tyrosinase (also called
tyrosine hydroxylase)
Alkaptonuria Homogentisic acid
oxidase
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Q 4.
a. Describe ketogenesis in liver. How are ketone bodies utilized in
extrahepatic tissues. (2+2)

KEY
(a)
Reactions of ketogenesis (structural formulas and
structure are not required)
Acetoacetyl-CoA Formation:
It is the starting material for ketogenesis, arises
either directly during the course of -oxidation or
as a result of the condensation of acetyl-CoA
(catalyzed by thiolase).
HMGCoA formation:
Mitochondrial HMG CoA synthase combines a third
molecule of acetyl CoA with acetoacetyl CoA to
produce HMG CoA. HMG CoA synthase is a rate-
limiting enzyme for ketogeneis.
Formation of acetoacetate
HMG CoA is cleaved to produce acetoacetate and
acetyl CoA, as shown catalyzed by HMG CoA lyase.
Formation of 3-hydroxybutyrate
Acetoacetate can be reduced to form
3-hydroxybutyrate with NADH as the hydrogen
donor. This reaction is catalyzed by 3-
hydroxybutyrate dehydrogenase
Formation of acetone
Acetoacetate can also spontaneously be
decarboxylated in the blood to form acetone -a
volatile, biologically non-metabolized compound
that can be released in the breath.

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UTILIZATION OF KETONE BODIES
In the extraheptic tissues utilization involves the conversion of acetoacetate to
acetoacetyl-CoA succinyl-CoA and the enzyme succinyl-CoA-acetoacetate CoA
transferase (thiophorase).
Acetoacetate reacts with succinyl-CoA, and the CoA is transferred to form
acetoacetyl-CoA, leaving free succinate.
The acetoacetyl-CoA formed by these reactions is split to acetyl-CoA by thiolase
and oxidized in the citric acid cycle.
Extrahepatic tissues, including the brain but excluding cells lacking mitochondria
(for example, red blood cells), efficiently oxidize acetoacetate and
3-hydroxybutyrate in this manner. But liver cannot utilize ketone bodies as its
lacks thiophorase.





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Q 5.
a. Write down the reactions catalyzed by alanine aminotransferase,
arginase, creatine kinase, dopamine hydroxylase. (2)
b. Describe sources of ammonia in body. (2)
KEY
(a)
Alanine + alpha-ketoglutarate Pyruvate + Glutamate (catalyzed by alanine
aminotransferase)
Arginine Urea + Ornithine (catalyzed by arginase)
Creatine + ATP Creatine phosphate + ADP (catalyzed by creatine kinase)
Dopamine + O
2
Norepinephrine + H
2
O (Catalyzed by dopamine
hydroxylase requiring vitamin C)

(b)
1- Amino acids are quantitatively the most important source of ammonia,
because amino acids are deaminated to produce ammonia. Many tissues, but
particularly the liver, form ammonia from amino acids by the aminotransferase
and glutamate dehydrogenase reactions.
2- From glutamine: The kidneys form ammonia from glutamine by the action
of renal glutaminase. Most of this ammonia is excreted into the urine as NH
4
+
.
Ammonia is also obtained from the hydrolysis of glutamine by intestinal
glutaminase. The intestinal mucosal cells obtain glutamine either from the blood
or from digestion of dietary protein.
3- From bacterial action in the intestine: Ammonia is formed from urea by the
action of bacterial urease in the lumen of the intestine. This ammonia is
absorbed from the intestine by way of the portal vein and is almost
quantitatively removed by the liver via conversion to urea.
4- From amines: Amines obtained from the diet, and monoamines that serve
as hormones or neurotransmitters, give rise to ammoia by the action of amine
oxidase.
5. From purines and pyrimidines: In the catabolism of purines and
pyrimidines, amino groups attached to the rings are released as ammonia.

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Q 6.
a. What is the genetic code and write down its three characteristics.
(2)
b. What is the basic function of type I DNA topoisomerases?
Describe any two types of covalent modifications in protein
molecules following their synthesis. ( +1)

KEY
(a)
The genetic code: The genetic code is a dictionary that identifies the
correspondence between a sequence of nucleotide bases and a sequence of
amino acids. Each individual word in the code is composed of three nucleotide
bases. These genetic words are called codons.
Characteristics of the genetic code
1. Specificity: The genetic code is specific (unambiguous), that is, a specific
codon always codes for the same amino acid.

2. Universality: The genetic code is virtually universal, that is, the specificity
of the genetic code has been conserved from very early stages of evolution,
with only slight differences in the manner in which the code is translated.
3. Redundancy: The genetic code is redundant (sometimes called
degenerate). Although each codon corresponds to a single amino acid, a
given amino acid may have more than one triplet coding for it. For example,
arginine is specified by six different codons.
4. Nonoverlapping and commaless: The genetic code is nonoverlapping
and commaless, that is, the code is read from a fixed starting point as a
continuous sequence of bases, taken three at a time.

(b)
Type I DNA topoisomerases: These enzymes reversibly cut a single
strand of the double helical DNA molecule. They have both nuclease
(strand-cutting) and ligase (strand-resealing) activities. They do not
require ATP, but rather appear to store the energy from the phosphodiester
bond they cleave, reusing the energy to reseal the strand. Type I
topoisomerases relax negative supercoils in E. coli, and both negative and
positive supercoils in eukaryotic cells.
Covalent Modifications in Proteins (Any two of the following
modifications)
1. Phosphorylation: Phosphorylation occurs on the hydroxyl groups of
serine, threonine, or, less frequently, tyrosine residues in a protein. This
phosphorylation is catalyzed by protein kinases.

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2. Glycosylation: Many of the proteins that are destined to become part of
a plasma membrane or lysosome or to be secreted from the cell have
carbohydrate chains attached to serine or threonine hydroxyl groups
(O-linked) or the amide nitrogen of asparagine (N-linked). The stepwise
addition of sugars occurs in the endoplasmic reticulum and the Golgi
apparatus.
3. Hydroxylation: Proline and lysine residues of chains of collagen are
extensively hydroxylated in the endoplasmic reticulum.
4. Carboxylation: Gamma-carboxylation of glutamate residues of various
clotting and other proteins is brought about by vitamin K The resulting
gamma-carboxyglutamate residues are essential for the activity of these
clotting and other proteins.
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Q 7.
a. How does parathyroid hormone tend to increase plasma calcium
concentration? (2)
b. Name the second messengers of atrial natriuretic peptide, ACTH.
Write down the steps for the synthesis of catecholamines from
tyorosine. ( +2)
KEY
(a)
PTH has the following ways to increase Ca
2+
level in serum
1. increases the rate of dissolution of bone by stimulating the osteoclastic
activity in bone, including both organic and inorganic phases, which moves
Ca
2+
into extracellular fluid;
2. reduces the renal clearance or excretion of calcium hence there is decreased
loss of calcium through kidney.
3. increases the efficiency of calcium absorption from the intestine by promoting
the synthesis of calcitriol as the activity of renal alpha1-hydroxylase is
increased due to the action of parathyroid hormone. It is the calcitriol that
causes the increased absorption of dietary calcium by inducing the synthesis
of calcium binding protein in the enterocytes.
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(b)
Second messengers
Atrial natriuretic peptide: Cyclic GMP (cGMP)
ACTH: Cyclic AMP (cAMP)
Steps for the synthesis of catecholamines
1- RING HYDROXYLATION OF TYROSINE TO L-DOPA: Tyrosine hydorxylase
coverts L-tyrosine into L-dopa (dihydroxyphenylalanine).
2- DECARBOXYLATION TO DOPAMINE: Dopa decarboxylase a pyridoxal
phosphate-dependent enzyme causes the conversion of L -dopa to dopamine.
3- SIDE-CHAIN HYDROXYLATION OF DOPAMINE TO NOREPINEPHRINE:
Dopamine -hydroxylase (DBH) converts dopamine to norepinephrine.
4- N-METHYLATION OF NOREPINEPHRINE TO EPINEPHRINE: PNMT
(phenylethanolmin-N-methyltransferase (PNMT) converts norepinephrine to
epinephrine.
The following structural formulas are not required

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Q 8.
a. What do you understand by balanced diet and basal energy
expenditure? Enumerate any six factors affecting basal energy
expenditure. (1+1)
b. Describe the buffering action of phosphate buffer system in
kidney. (2)

KEY
(a)
Balanced diet: It is the diet that contains all types of basic dietary
ingredients/components namely proteins, lipids, carbohydrates, vitamins,
minerals , and water, in quantities sufficient for the particular individual
depending upon his/her age, sex, etc.
Basal energy expenditure (BEE, also called basal metabolic rate or
BMR): It is the rate of energy expenditure under certain empirical
circumstances known as basal conditions and is expressed as kilocalories per
square meter of body surface area per hour.
Factors affecting basal energy expenditure: (Factors maybe other than the
following)
1. Age of the individual
2. Gender
3. Climate
4. Body temperature
5. Race
6. Diet
7. Hormones
8. Sleep
9. Pregnancy and lactation
10.Habits
11.Drugs
Buffering action of phosphate buffer system in kidney
Phosphate buffer system comprises H2PO4
2-
(proton donor) and HPO4
-
(proton
acceptor). It plays a major role in buffering the renal tubular fluid for the
following two reasons
1. phosphate usually becomes greatly concentrated in the tubules, thereby
increasing the buffering power of the phosphate system
2. the tubular fluid usually has a considerably lower pH bringing the operating
range of the buffer closer to the pK (6.8) of the system (as a buffer system
exhibits its maximum buffering action at pH near to its pKa).
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The excess of hydrogen ions are excreted into the tubular fluid combines with
the proton acceptor component of phosphate buffer system (HPO4
2-
) to form
H2PO
4
-
that is excreted in the form of sodium salt NaH
2
PO
4
in the urine.
Na
2
HPO
4
+ H
+
NaH
2
PO
4

The following figure is not required