M.

Prasad Naidu
MSc Medical Biochemistry, Ph.D,.

Peptides are short sequences of amino acids held by
peptide bonds.

Each peptide chain will have 2 ends an amino terminal [N]
and a carboxy terminal [C].

Peptide bonds are digested by peptidases and proteases.
1. Glutathione 3
2. TRH 3
3. Enkephalins 5
4. Angiotensin- II 8
5. Oxytocin 9
6. Vasopressin 9
7. Bradykinin 9
8. Aspartame 2

Glutathione (GSH)
Glutathione has a Pseudopeptide linkage.
GS-H is a Tripeptide 3 amino acids
It is gama glutamyl cysteinyl glycine
Glutathione is present in RBC and many other
tissues.
Reduced glutathione (GS-H) is the
active form



2G SH G - S S G
Reduced Oxidized
Active Inactive
Functions of Glutathione
1. Reduced glutathione is essential for maintaining the
normal structure of red blood cells.
Glutathione (reduced) performs specialized functions in
erythrocytes
It maintains RBC membrane structure and intergrity.
It protects hemoglobin from getting oxidized by agents
such as H
2
O
2
.
glutathione keeps iron in ferrous state in haemoglobin
there by preventing formation of methaemoglobin.
RBCs with lowered level of reduced glutathione are more
susceptible to haemolysis.

2. Glutathione serves as a coenzyme fro certain
enzymes.
eg: Prostaglandin PGE
2
synthetase.

3. It is essential for the formation of correct
disulfide bonds in several proteins.

4. Toxic amounts of peroxides and free radicals
produced in the cells are scavanged by glutathione
peroxidase (selenium containing enzyme)

Peroxidase
2GSH + H
2
O
2
G S S G - + 2H
2
O



5. Glutathione is involved in the transport of amino acids
in the intestine and kidney tubules via -glutamyl cycle or
meister cycle .

6. It keeps the enzymes in an active state by preventing
the oxidation of sulfhydryl (-SH-) group of enzyme to
disulfide (-s-s-) group.

7. As a conjugating agent in detoxification
(liver)






Conjugation for detoxification:

Glutathione helps to detoxify several compounds by
transfering the cysteinyl group

e.g:
a. Organo phosphorus compounds
b. Halogenated compounds
c. Nitrogenous substances ( chloro dinitro benzene)
d. Heavy metals
e. Drugs.
The reaction is catalyzed by glutathione- s
transferase (GST)


TRH - Thyrotropin-releasing hormone
Secreted by hypothalamus,

causes anterior pituitary gland to release
thyrotropic hormone

Methionine enkephalin - Enkephalins (5 Aas)

Opiate-like peptide.
Found in brain.
Inhibits sense of pain.

Angiotensin- II (8 AAs)

Pressor or hypertensive peptide
Stimulates release of aldosterone from
adrenal cortex.

Oxytocin (9 AAs)

Oxytocin secreted by posterior pituitary gland

Contains 9 amino acids(nonapeptide)
Oxytocin causes contraction of uterus.

Vasopressin (antidiuretic hormone) (9 Aas)

Secreted by posterior pituitary gland
causes kidney to retain water from urine

Plasma bradykinin Bradykinin 9 AAs

Vasodilator peptide
Produced from plasma proteins by snake
venom enzymes.
Aspartame -2AAs

Its a dipeptide produced commercially by
combination of aspartic acid and
phenylalanine.

It is above 200 times sweeter than sucrose
It is used as a low calorie artificial sweetner in
soft drink industry
Iso electric PH (PI) of an amino acid / protein
is the PH at which it has both + Ve and Ve
charges in equal quantities and as a whole it
is electrically neutral. Hence it does not move
in an electric field .protein precipitated.
Iso electric PH (PI) = PK 1 + PK 2
(for Monoamino, Monocarboxylic AA) 2
Ex : for glycine = 2.4 + 9.8 = 6.1
2
Isoelectric PH

Amino acids / Proteins are ampholytes - contain both
acidic (-COOH) and basic group(-NH
2
)

They can donate proton and accept a proton. Hence
they are ampholytes.

Zwitterion or Dipolar ion

Amino acids / Proteins act as zwitter ion (dipolar
ions) containing positive and negative ionic groups.

All the ionisable groups present in the protein will
influence the pI of the protein.

At isoelectric pH proteins will not migrate in
electric field .

Proteins have minimum solubility.
Hence easily precipitated.
Proteins have minimum buffering capacity .
Proteins have low viscosity.


From the graph it is evident that the
buffering action is maximum in and
around pK1 or at pK2 and minimum at pl
Amino acids can release H+ and act as
weak acids.
It can be quantitatively described by
Henderson Hasselbalch equation :
pH = pKa + log [A-] (Conjugate base)
[HA] (Acid)
It predicts maximum buffering occurs 1 pH around pKa
Proteins
Lysozyme
Pancreatic ribonuclease
Myoglobin
Hemoglobin
Human serum immunoglobulins
Carboxypeptidase
Casein
Fibrinogen
Human serum albumin
Egg albumin
Pepsin
PI
11
9.6
7.0
6.8
6.4 7.2
6.0
4.6
5.5
4.8
4.6
1.0
Isoelectric pH of some proteins
The pI value is characteristic for each protein. In a
solution at a pH value above its pI, a protein will have a
net negative charge(Anion) below its pI, it has a
positive charge(cation).

By this technique proteins are seperated (immobilized)
at Isoelectric pH during electrophoresis.

As the electrophoresis occurs proteins migrate to
positions corresponding to isoelectric pH.
Serum proteins can be seperated by to 40 different
bands.

Curdling of milk lactic acid formed during curdling of
milk brings the pH of milk to 4.6(Iso electric pH of
casein) where casein is precipitated.

Heat and Acetic acid Test;
Acetic acid is added to urine to bring the pH to around 4.8
(Iso electric pH of albumin) and then heated to detect
albumin in urine.



Non Protein Amino Acids
or
Non standard Amino Acids

Amino acids Functions
Amino acids
Ornithine
Citrulline Intermediates in the biosynthesis of urea
Arginosuccinic acid


Thyroxine
Triiodothyronine Thyroid hormones derived from tyrosine.


S-Adenosylmethionine Methyldonor in biological system.
Homocysteine Intermediate in methioninen metabolism.
A risk factor for coronary heart diseases.

Homoserine Intermediate in threonine, aspartate and
methionine metabolism.

3,4-Dihhydroxy phenylanine
(DOPA) A neurotransmitter, serves as a
precursor of melanin pigment.
Non standard amino acids
Non--amino acids
-Alanine Component of vitamin
pantothenic acid and coenzyme A

-Aminobutyric acid A neurotransmitter produced
(GABA) from glutamic acid

- Aminolevulinic acid Intermediate in the synthesis
(ALA) of porphyrin (finally heme)

Taurine Found in association with bile acids.