Protein metabolism

BY

Dr. NAGLAA IBRAHIM AZAB

Lecturer Of Medical Biochemistry

&Molecular Biology

BENHA FACULTY OF MEDICINE

 Protein structure
α- a.a ---- α- a.a ---- α- a.a ---- α- a.a ---------- α- a.a
1 2 3 4 n

Peptide linkages

α- amino acid

R—CH—COOH

NH2

R1 R2 Rn

NH2 -- CH—CO---- NH-- CH—CO----------CO---- NH- CH—COOH

Amino terminus peptide linkages carboxyl terminus

 Classification of amino acids
• Structural classification : according to •
the chemical structure of the side chain
(R)
• Nutritional classification •
(essensial & non essential a.as)
• Metabolic classification : according to •
the fate of amino acids inside the body
(glucogenic,ketogenic and mixed a.as)
 Classification
According to
R

Aliphatic a.as Heterocyclic a.as

Aromatic a.as

Contain heterocyclic
No ring structure ring
contain Phenyl ring
 Aliphatic a.as
Neutral a.as Amidic a.as
Contain Contain

One NH2 one COOH One NH2 one COOH

One NH2CO(amidic group)

Acidic a.as Basic a.as

Contain Contain

One NH2 two COOH >One NH2 one COOH

 Aliphatic neutral a.as
WITH HYDROCARBON SIDE CHAIN
--------------------------------------------------
1-Glycine(2C) CH2–
COOH I
NH2

2-Alanine(3C) CH3–CH–COOH
I
NH2

3-Valine(5C) CH3–CH–CH–COOH
I I
CH3NH2

4-Leucine(6C) CH3–CH–CH2–CH–COOH
I I
CH3 NH2

5-Isoleucine(6C) CH3–CH2–CH–CH–COOH
I I
CH3 NH2
WITH HYDROXYL (OH ) CONTAINING SIDE CHAIN
-----------------------------------------------------------------------

1-Serine(3C) CH2–CH–COOH
I I
OH NH2

2-Homoserine(4C) CH2–CH2–CH–COOH
I I
OH NH2

3-Threonine(4C) CH3–CH–CH–COOH
I I
OH NH2
 WITH SULFUR (S) CONTAINING SIDE CHAIN
---------------------------------------------------------------

1-Cysteine(3C) CH2–CH–COOH
I I
SH NH2
2-Homocysteine(4C) CH2–CH2–CH–COOH
I
SH

3-Methionine(4C) CH2–CH2–CH–COOH
I I
S-CH3 NH2
4-Cystine CH2–CH–COOH
S I
NH2
S
CH2–CH–COOH
I
NH2
 Aliphatic Aliphatic
acidic a.as Amidic a.as
Aspartic acid (4C) Aspargine (4C)

HOOC–CH2–CH–COOH NH2–OC–CH2–CH–
I COOH I
NH2 NH2

Glutamic acid (5C) Glutamine (5C)

HOOC–CH2–CH2–CH–COOH NH2–OC–CH2–CH2–CH–COOH
I I
NH2 NH2
 Aliphatic Basic a.as
1-Lysine (6C) CH2–CH2–CH2–CH2–CH–COOH
I I
NH2 NH2
2-Arginine (5C+1) CH2–CH2–CH2–CH–COOH
NH I
Guanido group I NH2
NH =C─ NH2

3-Ornithine (5C) CH2–CH2–CH2–CH–COOH

I I
NH2 NH2
4-Citrulline (5C+1) CH2–CH2–CH2–CH–COOH
I
NH
NH2
I
Ureido group
O= C─ NH2

Ornithine and citrulline are not found in proteins but are formed in urea cycle
 Aromatic a.as
Phenylalanine CH2–CH–COOH
I
NH2

Tyrosine = Parahydroxy phenylalanine

CH2–CH–COOH
I
NH2

OH
 Heterocyclic a.as
Tryptophan CH2–CH–COOH
I
N
NH2
H

Histidine CH2–CH–COOH
N NH
I
NH2

Proline (imino acid)

COOH
N

H
 .

Proteins

Parietal cells Gastric HCL

1st
Chief cells HOW? Denatured proteins

Pepsinogen Pepsin Endopeptidase with broad

Specificity (cleaves the
Inactive zymogen Active enzyme internal peptide bonds in
then which the carboxylic group
is of aromatic a.as or leucine)
Rennin in infants

Milk Ca Ca paracaseinate
Soluble casein
caseinogen (milk clot)
Denaturation
Importance?

Further digested by pepsin

and other proteases
Large peptides + some free a.as
 Large peptides + some free a.as

Pancreatic endopeptidases including

Intestinal cells
enteropeptidase
*Trypsinogen Trypsin Cleaves peptide bonds in which the COOH
(inactive) (active) is of Basic a.as (arginine and lysine)

*Chymotrypsinogen Chymotrypsin Cleaves peptide bonds in which the COOH

(inactive) (active) is of Aromatic a.as or leucine.

*Proelastase Elastase Cleaves peptide bonds in which the COOH

(inactive) (active) is of small non polar a.as (a.as with small
uncharged R as glycine, alanine & serine)

*Collagenase Catalyzes the hydrolysis of collagen

Smaller peptides + free a.as

 Exopeptidases
Smaller peptides + free a.as

Pancreatic carboxypeptidases including

*Procarboxypeptidase A Carboxy peptidase A Cleaves the aromatic a.as from the

(inactive) (active) C- terminal end of the peptide
Trypsin
*Procarboxypeptidase B Carboxy peptidase B Cleaves the basic a.as from the
(inactive) (active)
C- terminal end of the peptide

Intestinal proteases

*Aminopeptidases Cleaves one a.a from the

N- terminal end of the peptide

*Dipeptidases and tripeptidases Cleaves a.as from the dipeptides

or tripeptides

a.as
In infants, Ig A in the clostrum of milk is
absorbed without digestion by
pinocytosis
giving immunity to the babies

Intestinal lumen Intestinal cell Blood

IgA [gA [gA [gA IgA

IgA IgA
Aa.s resulting from protein digestion
are absorbed from the small intestine by:

■ passive transport mechanism (For D-aas).

■ Active transport mechanism (For L-aas and

dipeptides):
► Carrier protein transport system
( sodium – amino acid carrier system ).
► Glutathione transport system
(γ-glutamyl cycle)
 Carrier protein transport system
( sodium – amino acid carrier system )
Intestinal lumen
K+ Na+ a.a.

ADP+Pi Na+ a.a.

Cell membrane
Na/K ATPase
Carrier
ATP

Intestinal cell K+ Na+ a.a.

Cytoplasm

Portal blood a.a.

*This system transport the a.a. against its conc.
gradiant using energy derived from Na/K+ pump.
*Here a.as are absorbed by specific carrier protein
in the cell membrane of the small intestinal
cells.This carrier protein has one site for the a.a.
and another site for the Na+. It transports them
from the intestinal lumen across the cell
membrane to the cytoplasm. Then the a.a.
passes to the blood down its conc. Gradient,
while the Na+ is pumped out from the cell to the
intestinal lumen by Na/K+ pump utilizing ATP as a
source of energy derived from Na/K+ pump.
 Glutathione transport system
(γ-glutamyl cycle)
R
NH2—CH—COOHa.a.
γ-glutamyl transpeptidase(transferase) (GGT)
Cell membrane

CO-NH-CH-COOH
CH2 R
γ-glutamyl- cysteinylglycine cysteinylglycine CH2
CH-NH2
ADP+Pi
glutathione dipeptidase COOH
synthetase
ATP
glycine γ-glutamyl a.a.

γ-glutamyl- cysteine
γ-Glutamyl
a.a.
cyclotransferase
ADP+Pi γ-Glutamylcysteine
synthetase
cysteine
ATP 5-oxoproline

Glutamic acid
Oxoprolinase H2C CH2

H2O
O=C CH-COOH
ADP+Pi ATP N
I
H
►This transport system is for:
the transport of a.a.s from the extracellular space to the
cytoplasm in the intestine,kidney, brain & liver(bile ductule cells)

So it is not important only for the uptake of a.as from the

intestinal lumen (a.a. absorption) .

► 3 ATP molecules are utilized for the transfer of one

a.a.

►Clinical notes:
■The blood conc. of GGT enzyme is increased in
cholestasis & chronic alcholism ( so used as a liver
function test).
■Oxoprolinuria: Inherited deficiency of glutathione
synthetase enzyme , leading to increase levels of 5
oxoproline in blood &urine acidosis &
neurological damage
 Quiz
The basis of allergic reactions to
food
 Fate of absorbed a.as

Enter in the formation of a.a. pool

► Definition: It include the free a.as distributed throughout the body
- The a.a. pool contains 100 gm a.as.50% of these a.as are in the form of
glutamate & glutamine (Why?)
- In contrast to the amount of protein in the body (about 12 Kg in 70 Kg man),
the a.a. pool is small (only 100 gm)
►Sources & fate of the a.a. pool:
Non essential a.a.s
Diatery proteins Tissue proteins
synthesized in the body
a.a.s

Amino acid pool

Anabolism Catabolism(Deamination)
Synthesis of Fate of deamination products

Proteins Other nitrogenous compounds α- keto acid Ammonia

Krebs
►Aminosugars
►Tissue proteins cycle
►Nitrogenous bases
►Plasma proteins of phospholipids
►Enzymes Synthetic Catabolic
►Purines & pyrimidines glucose Ketone bodies CO2+H2O
►Some hormones +ENERGY pathway pathway
►Neurotransmitters
►Milk ►Niacin
►Creatine
►Heme Non essential
glutamine Excreted
a.a.synthesis Urea
in urine
 Catabolism of the a.a.s occurs by
Deamination (removal of the amino group)
either by
General methods of deamination:
►Oxidative deamination
►Transamination
►Transdeamination
Or
Specific methods of deamination:
For certain a.a.s
 Oxidative deamination
► Definition: It is the
oxidation (removal of hydrogen)
and deamination (removal of the amino group
which is liberated as free ammonia)
giving
α- ketoacid and ammonia (reversible reactions)

R— CH — COOH
l
NH2
► Site: In most tissues , mostly in the liver
and kidney
► Enzymes involved:
1- L-glutamate dehydrogenase enzyme:
Present in the cytosol & mitochondria of most tissues

NAD(P) NAD(P)H+H

H2O NH3

HOOC–CH2–CH2–CH–COOH ◄ ► HOOC–CH2–CH2–C –COOH

l L-GLUTAMATE ll
DEHYDROGENASE ENZYME
NH2 O

L-glutamic acid α- ketoglutaric acid

Coenzyme is NAD or NADP

Regulation: The direction of the reaction depends on:
1- Availability of the substrates:
--Relative conc. Of (α-ketoglutarate &NH3) and (glutamate).
--Ratio of NADP : NADPH+H
2- Allosteric regulation:
--Activators : ADP or GDP.
-- Inhibitors : ATP ,GTP & NADH
 QUIZ
---After a protein meal , in which direction
the reaction proceeds? Why?

---After a CHO meal ,

in which direction the
reaction proceeds?
Why?
2- D- & L- a.a. oxidases :
Present only in the liver and kidney in minimal amounts
They are of low activity in the mammalian tissue
N.B. a.a.s L-a.as : mammalian proteins are formed of only L-a.as
D-a.as: are found in plants and the cell wall of
microorganisms but not used in the synthesis of mammalian proteins

L-a.a. oxidase : It deaminates most of the naturally occuring a.a.s.

H2O L-a.a. oxidase
NH3
R— CH — COOH R— C — COOH
l ll
NH2 O
FMN FMNH2
L- amino acid α- keto acid
H2O2 O2
H2O2 Catalase

2 H2O + O2
D-a.a. oxidase : It deaminates D-a.as present in diet
giving α- keto acids
that either transaminated to the coressponding L-a.as
or converted to glucose or F.as
or catabolized to CO2 + H2O + energy

H2O NH3
D-a.a. oxidase
R— CH — COOH R— C — COOH
l ll
NH2 O
D- amino acid FAD FADH2 α- keto acid
L- amino acid
H2O2 O2
Transaminase
H2O2 Catalase α- keto acid

2 H2O + O2 ■Glucose
R— CH — COOH ■f.as
l
■CO2+H2O
NH2

L- amino acid
►Importance of oxidative deamination

L-glutamate dehydrogenase enzyme is the only a.a.

that undergoes oxidative deamination in the
mammalian tissue.

Oxidative deamination by L- glutamate dehydrogenase

is an essential component of transdeamination.
So it is important in deamination of most a.as.
 Transamination
►Definition: It is the transfer of amino group
from one α- a.a. to α- keto acid
to form a new α- a.a & a new α- keto acid
(reversible reaction)
► Enzymes involved: Transaminases or aminotransferases
Coenzyme: PLP (Pyridoxal phosphate)
R— C H — COOH R— C — COOH
l ll
NH2 O
◄ ►
Transaminase
PLP

R— C — COOH
◄ ►R— C H — COOH
ll l
O NH2
►Site: In the cytosol or both the cytosol & the
mitochondria of most cells especially the
liver

► All a.as except

threonine,lysine,proline&hydroxy proline
may undergo transamination
► α-ketoglutarate & glutamate are often involved in
transamination reactions

α- a.a. ◄
α- keto glutarate
►

Glutamate transaminase

◄ ►
α- keto acid Glutamic acid
► Transaminases of clinical importance are:
■ Alanine transaminase(ALT) =
{ Glutamate pyruvate transaminase(GPT)}:

Glutamic acid + Pyruvic acid ALT

PLP
Alanine + α- keto glutarate
CH3— C — COOH CH3— C H — COOH
ll l
O NH2

■ Aspartate transaminase(AST)=
{ Glutamate oxaloacetate transaminase(GOT)}:
ALT

Glutamic acid + Oxaloacetic acid PLP

Aspartic acid + α- keto glutarate

HOOC–CH2–CH–COOH
HOOC–CH2–C–COOH
l
ll
NH2
O
►Value of transamination:
■Function:
1- Degradation of a.as to form α- keto acids.
2- Synthesis of non essential a.as from
CHO.
■Diagnostic value:
Transaminases are normally intracellular enzymes.
They are elevated in the blood when damage to the
cells producing these enzymes occurs.
** Increase level of both ALT & AST indicates
possible damage to the liver cells.
** Increase level of AST alone suggest damage to
heart muscle ,skeletal muscle or kidney.
 Transdeamination
►Definition:
It is the combination of transamination & oxidative deamination.
It includes the transamination of most a.as with α– keto
glutarate to form glutamate then the glutamate is oxidatively
deaminated reforming α– keto glutarate and giving ammonia.
This provides a pathway by which the amino group of most
a.as is released in the form of ammonia.

α- a.a. ◄ ► α- keto glutarate ◄

► NH3
►NAD(P)H+H
L-GLUTAMATE
Glutamate transaminase DEHYDROGENASE ENZYME NAD(P)

► H2O
α- keto acid ◄ ► Glutamic acid ◄
 On biochemical basis explain
■Removal of ammonia from a.as can not be explained alone
by transamination nor by oxidative deamination alone
It can not be explained by transamination alone as no free
ammonia is liberated nor by oxidative deamination alone
as oxid. Deamination works efficiently only on glutamic
acid as L- glutamate dehydrogenase is of high activity in
the mammalian tissue,while the L- amino acid oxidase
which works on most a.as is of low activity.

■The formation of NH3 from α– a.as occurs mainly via the α–

amino nitrogen of glutamate
As this occurs by transdeamination, and L- glutamate is the
only a.a that undergoes oxidative deamination at an
appreciable rate in the mammalian tissue.
Another importance of transdeamination is we
can form α– a.a. from ammonia

α- a.a. ◄ ► α- keto glutarate ◄

► NH3
►NAD(P)H+H
L-GLUTAMATE
Glutamate transaminase DEHYDROGENASE ENZYME NAD(P)

► H2O
α- keto acid ◄ ► Glutamic acid ◄
 Specific methods of deamination
1- L- glutamate dehydrogenase:

Said before
2- Glycine oxidase:
as the mechanism of action of D- amino acid oxidase

H2O NH3
Glycine oxidase
CH2 — COOH CH — COOH
l ll
NH2 O
FAD FADH2
Glycine Glyoxylic acid
H2O2 O2
H2O2 Catalase

2 H2O + O2
3- Glycine cleavage system:

NAD NADH+H

CH2 – COOH + FH4 CH2─FH4 + NH3 + CO2

|
NH2

Tetrahydrofolic Methylene
Glycine
acid tetrahydrofolic
acid
4- Histidase: ( Non oxidative deamination)

Histidase – CH = CH – COOH
– CH2 – CH – COOH
N NH
|
NH3 N NH
NH2

Histidine Urocanic acid

5-Dehydratases: (Non oxidative deamination)
For hydroxy containing a.as (serine& threonine)

CH2–CH–COOH Serine CH2=C–COOH CH3–C–COOH

l l dehydratase l ll
OH NH2 PLP NH2 NH
Serine H2O α–aminoacrylic acid α–iminopropionic acid

H2O Serine
dehydratase
NH3
CH3–C–COOH
ll
O
Pyruvic acid
CH3–CH–CH–COOH CH3–CH2–C–COOH
l l Threonine ll
dehydratase
OH NH2 O
PLP
Threonine NH3 α–ketobutyric acid
6-Desulfhydrases:
The a.a. cysteine is deaminated by cysteine desulfhydrase

CH2–CH–COOH Cysteine CH2=C–COOH CH3–C–COOH

l l desulfhydrase l ll
SH NH2 PLP NH2 NH
Cysteine H2S α–aminoacrylic acid α–iminopropionic acid\

H2O
Cysteine
desulfhydrase
NH3
CH3–C–COOH
ll
O
Pyruvic acid
7-Hydrolytic deaminases: Deamination by H2O

Glutaminase & asparginase which catalyze the

hydrolytic deamination of glutamine & aspargine
respectively.
H2O NH3
OC–CH2–CH2–CH–COOH HOOC–CH2–CH2–CH–COOH
l l Glutaminase l
NH2 NH2 (in kidney, intestine) NH2

Glutamine Glutamic acid

8-Reductive deaminases:
By the action of intestinal bacteria on the a.as
(putrefaction) with the production of the
corresponding organic acids.

R— C H — COOH 2H NH3 R— C H2 — COOH

l
NH2

α- a.a. Corresponding organic acid

 Deamination products

NH3 α- keto acid

Blood level: Normally < 0.1 mg / dl

Urine level: 0.7 gm / day

Sources & fate:

 Various nitrogenous compounds
Amino acids
as

Purines& pyrimidines Some

(amino groups attached neurotransmitters Urea secreted into
Deamination: to the rings) as the intestine
--Oxidative deamination
--Transdeamination Monoamines:
--Specific deamination --serotonin
Methods
Histamine
Catabolism –epinephrine,norepinephrine,
( MAIN SOURCE) dopamine & their metabolites
metanephrine, normetanephrine Histaminase
&3 methoxy tyramine Intestinal bacterial
urease
Monoamine oxidase
(MAO)

NH3
Synthetic Catabolic
pathway pathway
Liver

Transdeamination (MAIN FATE)

Non essential glutamine Excreted

Urea
a.a.synthesis in urine
Urine
►Urea formation: is the main pathway by which the body gets rid of
NH3
►Glutamine formation:
--By the glutamine synthetase enzyme which is a mitochondrial enzyme
glutamine synthetase
Glutamic acid + ammonia glutamine

-- Glutamine is produced in many extra renal tissues esp. important:

•In the muscle
•In the liver:The formation of glutamine can be considered as a
mechanism for scavenging NH3 that has not been incorporated into
urea.
•In the brain: It removes the toxic effect of NH3 in the brain .Then the
glutamine goes via the blood to the kidneys where it become
hydrolyzed by glutaminase into glutamic acid and NH3 which is
excreted in urine,(This accounts for 60% of the NH3 excreted in urine)
glutaminase
Glutamine Glutamic acid

H2O NH3
►NH3 produced from a.a. deamination in the kidney is directly
excreted in urine ( This acconts for 40%of NH3 excreted in urine)

■N.B. NH3 produced from a.a deamination in the kidney esp.

glutamine regulates acid base balance &preserve cations
 Steps CO2 NH3 Cytoplasm

Mitochondria
CO2 + NH3
2ATP
Carbamoyl-phosphate synthetase 1
2 ADP+1 Pi
O O
ll ll
H2N – C – O – P – OH Carbamoyl-phosphate
l CH2–CH2–CH2–CH–COOH
OH
Pi
l l
CH2–CH2–CH2–CH–COOH Ornithine NH NH2
l l transcarbamoylase l
Ornithine Citrulline
NH2 NH2 O=C–NH2

HOOC–CH2–CH–COOH
O Ornithine Citrulline l
ll NH2
Aspartic acid
H2N–C–NH2 Urea
Arginase Arginino-succinate ATP
synthetase

CH2–CH2–CH2–CH–COOH AMP+PPi
l l H2O H2O
CH2–CH2–CH2–CH–COOH
NH NH2
l l
l Arginine Arginino succinate
NH NH2
NH2–C=NH
l
Arginino succinase NH–C=NH
l
HOOC–C–H Fumarate HOOC–CH2–CH–COOH
ll
H–C–COOH
 Carbamoyl- phosphate synthetase I is different from
carbamoyl phosphate synthetase II

CPS I CPS II
Site Mitochondria Cytoplasm

Pathway Urea synthesis Pyrimidine synthesis

+ve N-acetyl Nil

effector glutamate
Inhibitor Nil CTP

Source Ammonia Glutamine

of N
 O
Sources of the atoms of urea: ll
H2N–C–NH2

1- C & O: from CO2

2- 1st N atom: from NH3
3- 2nd N atom: from aspartate

Glutamate is usually the immediate precursor

of both NH3 &aspartate

H2O Glutamate
Oxaloacetate
NAD

NADH+H Oxidative α- ketoglutarate

α- ketoglutarate deamination Transamination

NH3 Aspartate
Overall reaction:

NH3 + CO2 + Aspartate Urea + fumarate

There in no net gain or loss of ornithine, citrulline ,

argininosuccinate or arginine.

Ornithine is regenerated with each turn of the urea cycle.

The release of the high energy phosphate of carbamoyl
phosphate as inorganic phosphate drives the reaction
in the forward direction.
 Fate of fumarate & its link to TCA cycle

CO2 NH3
Oxaloacetate

Carbamoyl-phosphate
Malate NADH+H
NAD
3ATP
Fumarate
Ornithine Citrulline TCA CYCLE

Ornithine Citrulline
Aspartate
Urea
Arginine Arginino succinate

Fumarate Transamination

Cytoplasmic H2O
fumarase
Malate or Fasting
dehydrogenase state
Malate Malate Oxaloacetate PEP Glucose
shuttle
Fed state
NAD NADH+H
NADP
Malic
NADPH+H enzyme
3ATP Oxaloacetate ATP
citrate
lyase
Pyruvic acid Pyruvic acid Citrate Citrate Acetyl COA f.a.
synthesis
Bioenergetics:
Urea cycle consumes four "high-energy" phosphate bonds (3 ATP hydrolyzed to 2
ADP and one AMP).
1 ATP ADP + Pi
~P
1 ATP ADP + Pi
Adenosine ~ P
1 ATP AMP + Pi + Pi
~P

►However. One NADH+H molecule is produced by oxidative deamination of

glutamate to NH3 and α-ketoglutarate. Glutamate provides the NH3 used in the
initial synthesis of carbamoyl phosphate.

► Also fumarate in the cycle may be converted to malate in the cytosol . Malate then
oxidized to oxaloacetate gives 1 NADH+H equivalent to 3 ATP obtained from
3ADP,
So the net energy expenditure is only one high energy phosphate .

The two NADH+H produced can provide energy for the formation of 5 ATP, a net
production of one high energy phosphate bond for the urea cycle. However, if
gluconeogenesis is underway in the cytosol, the latter reducing equivalent is used
to drive the reversal of the glyceraldehyde 3-p dehydrogenase step instead of
generating ATP. So the net energy expenditure is only one high energy phosphate
.
 Regulation:
Carbamoyl phosphate synthetase 1 is the key & it has an absolute requirement
for

N-Acetylglutamic acid which act as an allosteric activa tor.

The synthesis of N- Acetylglutamate from glutamic acid & acetyl CO A is ++ by

high protein diet & a.as especially arginine → ↑rate of urea cycle
. •

Acetate GLUTAMATE Acetyl CO A

a.as esp arginine

Synthetase ++

N- ACETYL GLUTAMATE CO A

ALLOSTERICALLY ++
carbamoyl phosphate synthetase 1
 Structure:
Creatine (methyl guanido acetic acid)

CH2 – COOH
l Creatinine
CH3 – N
l
CH2 – CO
HN = C – NH2
l
CH3 – N
l
Phosphocreatine (Creatine phosphate) HN = C ––– NH
CH2 – COOH
l
CH3 – N O
l ll
HN = C – NH ~ P– OH
l
OH
 Synthesis of creatine& creatine phosphate
& their degradation into creatinine
Creatine is formed from 3 a.as ( glycine, arginine & methionine )
Glycine
Arginine + Glycine transamidinase Guanidoacetic + Ornithine
CH2 – COOH
(kidney, pancreas) acid
CH2–CH2–CH2–CH–COOH
l l | CH2 – COOH CH2–CH2–CH2–CH–COOH
NH2 l l l
NH NH2
NH NH2 NH2
l
l
NH = C–NH2 HN = C – NH2

Methyl transferase
S- adenosyl methionine

(liver)
S- adenosyl homocysteine

Creatine phosphate Creatine phosphokinase Creatine

(methyl guanido acetic acid) CH2 – COOH
(liver, brain, muscle) l
CH2 – COOH CH3 – N
l Slow l
Rapid Non enzymatic (spontaneous)
CH3 – N O HN = C – NH2
l ll Pi H2O
HN = C – NH ~ P– OH
l Creatinine CH2 – C O
OH l
CH3 – N
l
Excreted in urine HN = C ––– N H
Importance of creatine:
►Creatine is converted to creatine phosphate using ATP as a phosphate donor. Creatine
phosphate acts as a store of high energy phosphate ,so it is used as a source of energy
in time of need (as it can give the phosphate group to ADP to form ATP).

►Creatine is present mainly in the muscle (98%) , but also in the brain & blood . Both the
muscle & the brain contain large amounts of creatine phosphate.

►During muscular relaxation,creatine becomes converted to creatine phosphate to be used

as a source of energy during muscular contraction.
(During ms contraction, ATP is mostly derived from glycolysis for which ms glycogen acts
as a substrate,However before ATP from glycolysis become available, phosphocreatine
acts as a temporary source of ATP.So it is especially important during the early stages
of ms exercise(1st few minutes),where the largest quantities of creatine –P are found ).

Level of creatine in the blood& urine:

►Plasma creatine level = 0.4 mg %
Whole blood creatine level = 2 - 4 mg %

► Normally little creatine is excreted in urine ( normally < 100 mg /day ) .This is more in
females & children due to low androgens which increase the uptake of creatine by ms.
 Creatinuria
►Def.: It is the presence of large amounts of creatine in urine
(Normal level of creatine excreted in urine is < 100 mg / day.)

►Causes:
1- Physiological creatinuria :
■ in children: due to - ↓androgens → ↓ uptake of creatine by the muscle
- small mass of the ms
■ in female: due to - ↓ free androgen especially during pregnancy.
- after labour due to involution of the uterus.

2- Pathological creatinuria :
■ in males with ↓ androgens( hypogonadism )
■ in muscle atrophy as in myopathies.
■ ↑ protein catabolism in --hormonal disturbances as D.M. ,
hyperthyrodism and cushing syndrome,--also in fever and
starvation
Creatinine levels in serum& urine:

►Normal serum creatinine level = 0.5 - 1.5 mg%.

↑ serum creatinine level

Causes: 1- Impairment of kidney function : about 50% of kidney function must be

lost before serum creatinine is ↑.
2- ↑ ms mass : as acromegally & gigantism.

↓ serum creatinine level

Causes: ↓ ms mass as in - myasthenia gravis

- muscular dystrophy
- debilitation
► Normal urine creatinine level = 1 - 1.5 mg %.

- It is more in male due to greater ms mass ---- male = 1.5 g / day.

---- female = 1 gm / day.
- There is a steady production of constant amount of creatinine that is
proportional to the total amount of phosphocreatine & creatine in the
body , which is in turn proportional to the ms mass of the individual .
So, the serum & urine creatinine is constant for an individual &
approximately proportional to the ms mass.

Creatinine excretion rate (creatinine coefficient) :

•It is the amount of creatinine measured in mg / kg body weight / day.

•It is said to be remarkably constant (21 in male & 16 in female).(Why?)

So, creatinine excretion rate (creatinine coeffecient) can be used to

check the
accuracy of 24 hr collection of urine. If found lower than expected, it
indicates that part of the urine was discarded .
 Creatinine clearance
- The volume of serum or plasma that would be cleared of creatinine by one minute's excretion of
urine. value that reflects the body's ability to excrete creatinine; it is used to diagnose and
monitor renal function.
- Urinary creatinine is 100% endogenous in origin i.e. not affected by diet being synthesized in
the body & no diurnal variation .So it can be used for calculation of urinary output of substance
amount of creatinine in urine.
- Also creatinine is chiefly filtered by the kidney, though a small amount is actively secreted.
There is little-to-no tubular reabsorption of creatinine. If the filtering of the kidney is deficient,
blood levels rise. As a result, creatinine levels in blood and urine may be used to calculate the
creatinine clearance (CrCl), which reflects the glomerular filtration rate (GFR). The GFR is
clinically important because it is a measurement of renal function. However, in cases of severe
renal dysfunction, the creatinine clearance rate will be "overestimated" because the active
secretion of creatinine will account for a larger fraction of the total creatinine cleared. Ketoacids,
cimetidine and trimethoprim reduce creatinine tubular secretion and therefore increase the
accuracy of the GFR estimate, particularly in severe renal dysfunction. (In the absence of
secretion, creatinine behaves like inulin.)
- Creatinine clearance (CCr) can be calculated if values for creatinine's urine concentration
(UCr), urine flow rate (V), and creatinine's plasma concentration (PCr) are known. Since the
product of urine concentration and urine flow rate yields creatine's excretion rate, creatinine
clearance is also said to be its excretion rate (UCr×V) divided by its plasma concentration. This
is commonly represented mathematically as :
Urinary creatinine conc.X urine volume in ml
- Creatinine clearance(ml/min)=
Plasma creatinine conc X time of collection/min
 NPN Compounds
Def: Are nitrogenous compounds not
precipitated by protein ppting
reagents.

Include:
a.as NH3
Urea Uric acid
Creatine Creatinine
 a.as Ammonia Urea

Structure R-CH – CooH NH3 O

‫׀‬ ‫׀׀‬
NH2 H2N – C- NH2
Source  Proteins in diet.  a.a. deamination (main  Co2.
 Tissue protein catabolism. source)  NH3.
 Synthesis of non essential a.as.  other nitrogonous  Aspartic acid
(sources of a.a.pool) compounds as: Through the
- purines & pyrimidmes. Urea cycle in liver.
- amines.
-urea splitted in the intestine.
Fate ■Synthesis of :  Synthesis of a.as. Excretion in urine &
 Proteins as:  Urea formation. part Pass to
- tissue Pr.  Glutamine formation. intestine.
- Plasma Pr.  Excretion in urine.
- Enzymes.
- Some hrs.
- Milk.
 Other nitrogenous comp as creatine,heme,
neurotransmitters, purines & pyrimidinet
■catabolism (deamination) giving:
- NH3  Fate said with NH3.
- α -Keto-acids:→ glucose.
→ ketone bodies.
→ Co2 + H2o energy.
Blood level 4-6 mg/dl < 0.1 mg/dl. 20-40 mg/dl
Urine level 0.7 g/day 0.7 gm/day 20-40 gm/day.
Clinical Amino acid uria Ammonia intoxication Bl. Urea is a krdnay
implication function test  in
renal failure.
 Creatine Creatinine Uric acid

Structure CH2 – COOH CH2 – C O

l l
CH3 – N CH3 – N
l l
HN = C – NH2 HN = C ––– N H

Source - Arginine + glycine + -Creatine & creatine phosphate. Catabolism of purines

methionine. ( in ms, brain and other tissues.) (liver).
( in kidneys and liver.)

Fate Uptake by ms and other Excretion in urine. Excretion in urine.

tissues.
Bl. Level 0.4 mg % plasma 0.5-1.5 mg% 2-7 mg/dl
2-4mg% blood
Urine level < 100 mg/day 1-1.5 gm/day 500 mg/day

Clinical Creatinuria  serum creatinine: Hyperuricemia

implications - Renal failure. (gout)
- ↑ms mass as acromegally & Hypouricemia.
gigantism.
 serum creatinine in  ms mass as
ms dystrophy
deblitation
Creatinine coefficient for accuracy
of collecting 24 hrs volume of urine.
Creatinine clearance as measure of
GFR.
 Heterocyclic a.as
1- Tryptophan
Synthesis: essential a.a.
Catabolism: Both glucogenic and ketogenic.
Tryptophan
Tryptophan pyrrolase (oxygenase)

N- formyl kynurenine
Acetoacetic acid
-keto adipic acidα
α-aminomuconate
α-aminomuconate semialdehyde
Quinolinic acid
NAD&NADP
Nicotinic acid
2-Acrolyl 3- amino fumarate
3-Hydroxyanthranilic acid
Hydroxykynurenine
Kynurenine

Importance: Synthesis of 1-nicotinic acid(niacin)→ NAD&NADP.

2-Serotonin 3-Melatonin
4-Indole &skatole
1-Nicotinic acid:
• It is produced from tryptophan in the liver since the enzyme picolinate carboxylase has low activity in the liver so some 2-acrolyl 3-
aminofumarate is cyclized in non enzymatic reaction to niacin. •
Every 60 mg tryptophan gives 1 mg niacin. 50%of niacin is formed from tryptophan while the rest are obtained from diet.
Pellagra:
•Causes : 1- ↓tryptophan due to deficiency in diet as in maize eating areas or↓ absorption in hartnup disease
2-niacin deficiencyin diet or due to decrease its synthesis from tryptophan in case of carcinoid syndrome .
3- Isoniazid drug administration in T.B.treatment leading to vitamin B6 deficiency which is important in formation
of niacin from tryptophan
• pellagra is characterized by 3Ds diarrhea,dermatitis and dementia