Metabolism of Proteins

Roshan Ali
Assistant Professor
IBMS, KMU
Course Contents
• Metabolism of Protein
• Review the digestion and absorption of Proteins
• Nitrogen balance
• General pathway of Protein metabolism
• De-amination
 Digestion
and
Absorption
of
Proteins

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 Digestion of dietary proteins
Digestion is the disintegration of complex nutrients into simple,
soluble and assimilable form.
Most of the nitrogen in the diet is consumed in the form of
proteins. Proteins are too large to be absorbed. The dietary
proteins are hydrolyzed to amino acids by proteolytic enzymes,
which can be easily absorbed.

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• Proteolytic enzymes responsible for degrading proteins are
produced by three different organs;

• stomach,
• pancreas
• small intestine.

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 Characteristics of proteolytic enzymes

1)The y are hydrolases.

2)Secreted in the zymogen (Inactive) form,
converted to active form in the intestinal lumen.
The active site of the enzyme is masked by a small region of the
peptide chain that is removed by hydrolysis of a specific peptide
bond.
3) They may be Exopeptidases or Endopeptidases. Exopeptidases
catalyze the hydrolysis of peptide bonds, one at a time, from the
ends of peptides. Endopeptidases hydrolyze peptide bonds between
specific amino acids throughout the molecule.

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 Digestion in the oral cavity

There are no proteolytic enzymes present in the saliva. The function of

the saliva is to lubricate the food, this helps in making food soluble for
the action of proteolytic enzymes.
After mastication and chewing, the bolus of food enters stomach where
it is acted upon by gastric juice.

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Digestion by gastric juice
Gastric juice contains

HCL
and 4 proteolytic enzymes:
Pepsin,
Rennin,
Gastriscin and
Gelatinase.

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 Functions of HCL
• Too dilute to hydrolyze but can cause denaturation of dietary
proteins
-
• Required for activation of inactive proteolytic enzymes
secreted by the gastric mucosa.
-
• Has a strong bacteriostatic action.

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 Functions of Proteolytic enzymes Pepsin

-Secreted in the form of pepsinogen, activated by

HCL and pepsin itself (Autocatalysis)
-Acid stable endopeptidase, optimum pH lies
between 1.6-2.5 ; becomes inactive if the pH is > 5.0
-Active for a peptide bond where amino group is
contributed by aromatic amino acids like Phenyl
Alanine, Tyrosine and Tryptophan.
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Biochemistry for Medics
 Functions of proteolytic enzymes
Rennin

-Present in infants but absent in adult gastric juice.

- Secreted as pro-rennin
-Optimum p H 4.0
-Action is similar to pepsin
-Acts on casein of milk and is involved in the curdling of milk
-Commercially, ‘rennet’ tablets are used in the making of
cheese.

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 Functions of proteolytic enzymes
Gastriscin and Gelatinase
-Both are secreted in zymogen form and are acid-
stable. Activated in the presence of HCL.

-Gelatinase acts on Gelatin to hydrolyze it to short

peptides.

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 Digestion by Pancreatic Juice

The bolus of food after leaving stomach reaches duodenum and is acted upon by the
pancreatic juice. All the enzymes are active only in the alkaline medium and alkalinity is
provided by Bile juice and bicarbonates present in pancreatic juice. The proteolytic enzymes
present in pancreatic juice are:
-Trypsin
-Chymotrypsin
-Elastase
-Collagenase
- Carboxypeptidases
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 Role of Trypsin

-Secreted in the zymogen form (Trypsinogen)and

activation is brought about by Enterokinase (secreted by
the intestinal mucosa ) and by Trypsin itself. Trypsin is
specific for cleaving peptide bonds contributed by basic
amino acids.
Required for activation of Chymotrypsin
 Required for conversion of pro-elastase to elastase
It is required for conversion of fibrinogen to fibrin
It has weak action on casein
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 Role of Chymotrypsin

-Activated by Trypsin and Chymotrypsin itself

(auto catalytically )
-Cleaves peptide bonds contributed by aromatic amino
acids.
- Can hydrolyze milk protein.

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 Role of Carboxy-peptidases

Two types of Carboxypeptidase are there

Carboxypeptidase A:Metalloenzyme , contains Zinc, Activation
brought about by Trypsin and auto catalytically by itself,
Exopeptidase, Hydrolyzes the terminal peptide bond connected
to an end amino acid (Aromatic) bearing free carboxyl group
Carboxypeptidase B: Also an Exopeptidase, hydrolyzes terminal
peptide bonds connected by basic amino acids

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 Role of Elastase and Collagenase
Elastase-
Activated by Trypsin
Has maximum activity on peptide bonds contributed by
carbonyl groups of neutral aliphatic amino acids- Alanine,
Glycine, Serine etc.

Collagenase- Acts on Collagen

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Digestion by intestinal Juice

Enzymes present in intestinal juice are:

- Enterokinase

- Amino peptidases

- Prolidase

- Di and Tri peptidases

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Functions of the Intestinal enzymes
Enterokinase - required for activation of Trypsin, also called
Enteropeptidase, present in the epithelial cells of brush border
of duodenal mucosa. Bile salts help in its liberation in to
intestinal lumen.
Amino peptidases – are exopeptidases, remove the amino acid
one by one from amino terminal end of peptide chains.

Prolidase - also an exopeptidase, removes proline residues

from terminal end of peptide chains.
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 Functions of Intestinal
enzyme(Contd.)
Di and Tripeptidases - Digestion of di and tri peptides is brought
about by di and tri peptidases present in brush border membrane
of epithelial cells as well as present in the interior of the cell.
Tripeptidases convert tripeptides into a dipeptide and a free amino
acid, then dipeptidases convert dipeptides to free amino acids.

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Absorption of amino acids

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 Absorption of amino acids
Proteins are completely digested to constituent amino acids.
But some amounts of oligopeptides may remain undigested.
The products of digestion are rapidly absorbed.
Site of Absorption- Oligopeptides are absorbed from
duodenum. and proximal Jejunum, while amino acids are
absorbed from ileum and distal jejunum .

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 Absorption of amino acids

Mechanism of Absorption-
The absorption takes place by active transport (same as that of glucose). Natural L-
amino acids are actively transported.
D- amino acids are absorbed by simple diffusion.
Vitamin B6 is involved in the active transfer of amino acids.
Energy requiring process, ATP is required as a source of energy
A carrier protein is also required which may be Na+ dependent or independent.
Different carrier proteins are there specific for different amino acids.

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 Absorption of Oligopeptides
Absorbed by active transport

Intracellular peptidases hydrolyze them to amino acids

Hydrolysis is rapid to keep peptide concentration low in the cell

Transport mechanism is independent of L- amino acids

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 Role of glutathione

Glutathione participates in the Active group translocation of L-

amino acids in to the cells of small intestine, kidneys, seminal
vesicles, epididymis and brain. A cyclic pathway, operates in
which Glutathione is regenerated again, it is named as Gamma
Glutamyl Cycle or Meister cycle( Based on the name of
Scientist).

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Transamination & Deamination of Amino Acids and urea cycle
 CONTENT
• TRANSDEAMINATION
• TRANSAMINATION
• DEAMINATION
 TRANSDEAMINATION
The amino group of amino acids is released
by a coupled reaction called
TRANSDEAMINATION

Transamination followed by (Cytoplasm of all

body cells)

oxidative deamination (Mitochondria of liver

cells)
 Transamination
• Transamination  is a chemical reaction between two molecules.

• One is an amino acid, which contains an amine (NH2) group.

• The other is a keto acid, which contains a keto (=O) group.

• In transamination, the NH2 group on one molecule is exchanged with the =O group on the
other molecule. The amino acid becomes a keto acid, and the keto acid becomes an amino
acid.

• Transamination in biochemistry is accomplished by enzymes called transaminases or

aminotransferases.
Reaction
General scheme of transamination
R CH COOH + HOOC C CH2CH2COOH
NH2 O
aminokyselina
amino acid 2-oxoglutarate
2-oxoglutarát

aminotransferase
aminotransferasa
pyridoxalfosfát
pyridoxal phosphate

R C COOH + HOOC CH CH2CH2COOH

O NH2

2-oxokyselina
2-oxo acid glutamát
glutamate
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 DEAMINATION
• Deamination is the removal of an amine group from a molecule. 

• Enzymes which catalyze this reaction are called deaminases.

• In the human body, deamination takes place primarily in the liver,

however glutamate is also deaminated in the kidneys.

• Deamination is the process by which amino acids are broken down if

there is an excess of protein intake. The amino group is removed from
the amino acid and converted to ammonia.
 Deamination of amino acids

•Deamination - elimination of amino group from amino acid

with ammonia formation.

• Four types of deamination:

• - oxidative (the most important for higher animals),

• - reduction,
• - hydrolytic, and
• - intramolecular
 Reduction deamination:

R-CH(NH2)-COOH + 2H+  R-CH2-COOH + NH3

amino acid fatty acid

Hydrolytic deamination:

R-CH(NH2)-COOH + H2O  R-CH(OH)-COOH +

NH3
amino acid hydroxyacid

Intramolecular deamination:

R-CH(NH2)-COOH  R-CH-CH-COOH + NH3

amino acid unsaturated fatty acid
 Oxidative deamination
 During oxidative deamination, an amino acid is converted into the
corresponding keto acid by the removal of the amine functional group
as ammonia.
 The amine functional group is replaced by the ketone group. The
ammonia eventually goes into the urea cycle.
 Oxidative deamination occurs primarily on glutamic acid because
glutamic acid was the end product of many transamination reactions.
 The glutamate dehydrogenase is controlled by ATP and ADP. ATP acts as
an inhibitor whereas ADP is an activator.
Intake, catabolism, and excretion of nitrogen
proteolysis
Proteins amino acids
transamination

glutamate
detoxication dehydrogenation + deamination
in other tissues
glutamine NH3
deamidation
in kidney detoxication in liver

glutamate + NH4+ urea

deamination (excretion by urine) (excretion by urine)
in kidney

2-oxoglutarate + NH4+
(excretion by urine)

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Introduction of Urea
• Discovered in 1727
• By Dutch scientist Herman Boerhaave
• In 1828, the German chemist Friedrich Wohler obtained urea artificial
• By treating silver cyanate with ammonium chloride.
AgNCO + NH4Cl → (NH2)2CO + AgCl
Introduction of Urea (cont.…)
• This was the first time an organic compound was artificially synthesized from
inorganic starting materials, without the involvement of living organisms.
• Wohler said
"I must tell you that I can make urea without the use of kidneys, either man or dog.
Ammonium cyanate is urea."
Introduction of Urea (cont.…)
• For this discovery, Wohler is considered by many the father of organic chemistry.
Urea Cycle
 Urea Cycle
• Also known as “ornithine cycle”

• cycle of biochemical reactions occurring in many animals

• Produces urea ((NH2)2CO) from ammonia (NH3).

• In mammals, the urea cycle takes place primarily in the liver, and to a lesser extent in
the kidney.
 Urea Cycle (cont.…)
• Urea is synthesized in the liver

• Than secreted into blood stream

• And taken up by the kidneys for excretion in the urine.

• Partly takes place in mitochondria and partly in cytoplasm.

• End product of protein metabolism.

 Urea Cycle (cont.…)
• Ammonia is toxic to body

• Detoxified and converted into urea in urea cycle.

• The overall reaction of the urea cycle is as follow:

NH4+ + HCO3- + H2O + 3ATP + Aspartate Urea + 2ADP + AMP +

2Pi + PPi + Fumarate
Thank You