Amino acids

Proteins of food
Metabolites of
glycolysis and
Krebs cycle
Anabolic ways
Catabolic ways
Synthesis of
cell and
extracell
proteins
Synthesis of
peptide
physiologi-
cally active
substances
Trans-
ami-
nation
Deami-
nation
Decar-
boxila-
tion
Urea, CO
2
, H
2
O
Amines
Proteins and peptides of
the organism
GENERAL PATHWAYS OF AMINO ACIDS METABOLISM
Digestion and absorbtion of proteins in
the gastrointestinal tract. Nitrogenous
balance. Pathways of amino acids
transformation in an organism. Ways
of free ammonia neutralization.
Synthesis of urea.
Nitrogenous balance
The ratio between the amounts of nitrogen entered the
organism and nitrogen removed from the organism is called
nitrogenous balance. It may be positive, negative and
neutral (zero).
Positive nitrogenous balance the amount of nitrogen
entered the organism is more than amount of nitrogen
removed from the organism. It occurs in young growing
organism, during recovering after severe diseases, at the
using of anabolic medicines.
Negative nitrogenous balance the amount of nitrogen
removed from the organism is more than amount of nitrogen
entered the organism. It occurs in senile age, destroying of
malignant tumor, vast combustions, poisoning by some
toxins.
Zero nitrogenous balance the amount of nitrogen removed
from the organism is equal to the amount of nitrogen entered
the organism. It occurs in healthy adult people.

























Essential and unessential amino acids.

Essential amino acids are those amino acids which are not
synthesized in organism and must come with food.
Unessential amino acids can be synthesized in
organism from another compounds. There are 10
essential amino acids:
arginine, histidine, isoleucine, leucine, lysine,
methionine, phenylalanine,
threonine, tryptophan and valine.
Protein function in the organism.
- All enzymes are proteins.
- Storing amino acids as nutrients and as building
blocks for the growing organism.
- Transport function (proteins transport fatty acids,
bilirubin, ions, hormones, some drugs etc.).
- Proteins are essential elements in contractile and
motile systems (actin, myosin).
- Protective or defensive function (fibrinogen,
antibodies).
- Some hormones are proteins (insulin,
omatotropin).
- Structural function (collagen, elastin).
Protein digestion in the small intestine.
Most of the digestion of proteins is carried
out in the small intestine by pancreatic
enzymes trypsin, chymotrypsin,
carboxypeptidases and aminopeptidases.
PH of intestinal juice is 7-8.
Proteolytic enzymes and their activation.
Three enzymes are in gastric juice: pepsin,
gastricsin and rennin. All these enzymes cleave
proteins or peptides.
The main enzyme is pepsin. It is an endopeptidase
that cleaves peptide bonds in the interior of protein
substrate. The optimal pH for pepsin activity
1,5-2. Thus, hydrochloric acid is necessary for
pepsin functioning.
Special cells in the gastric mucosa produce
pepsinogen. Pepsinogen is converted to pepsin in
the gastric cavity. For this process the ready
pepsin and hydrochloric acid are necessary
(autocatalysis). Optimal pH for gastricsin is 2,0-
3,0. The ratio between gastricsin and pepsin in
gastric juice is 1:5,5. This ratio can be changed in
some pathological states.
Rennin also possesses a proteolytic activity and
causes a rapid coagulation of ingested casein. But
this enzyme plays important role only in children
because the optimal pH for it is 5-6.
Chemical composition of digestive juices.
Gastric juice contains water, enzymes,
hydrochloric acid, mineral salts and other
compounds. About 2,5 l of gastric juice is secreted
per day.
Intestinal juice contains water, enzymes, bile,
mineral salts, some tissue hormones and other
compounds.
The role of hydrochloric acid in digestion.
Denaturate proteins (denaturated proteins easier
undergo digestion by pepsin than native proteins).
Stimulates the activity of pepsin.
Hydrochloric acid has bactericidial properties.
Stimulates the peristalsis.
Regulate the enzyme function of pancreas.

Protein digestion in the small intestine.

Most of the digestion of proteins is carried out in the small intestine by
pancreatic enzymes trypsin, chymotrypsin, carboxypeptidases and
aminopeptidases.
PH of intestinal juice is 7-8.
Trypsinogen is produced by pancreas. This is not active form of
enzyme. Another enzyme enterokinase is secreted from the mucosal
cells of the small intestine. The optimal pH for trypsin 7,8.
Chymotrypsin is also produced by pancreas in inactive form
(chymotrypsinogen).. Chymotrypsin like trypsin also is endopeptidase.
It makes deeper hydrolysis of protein and splits about 50 % of all
peptide bonds basically those bonds that are formed between aromatic
amino acids.

Peptides of different size and structure are
finally decompoused in small intestine by
peptidases. Peptidases splitting the amino
acid from the end of free COOH group are
called carboxipeptidases and those, which
split amino acid residue from the end of free
NH2 group are called aminopeptidases.
The splitting of elastin in an intestine is
catalyzed by elastase and collagen is
decomposed by collagenase.
Digestion of protein takes place not only in
the intestinal cavity but also on the surface of
mucosa cells.
Mechanism of amino acid absorbtion.
This explanation is called the sodium cotransport
theory for amino acid transport; it is also called secondary
active transport of amino acid.
Absorption of amino acids through the intestine mucosa can occur
far more rapidly than protein can be digested in the lumen of the intestine.
Since most protein digestion occurs in the upper small intestine most
protein absorption occurs in the duodenun and jejunum.










The sources of amino acids:
1) absorption in the intestine;
2) formation during the protein decomposition;
3) synthesis from the carbohydrates and lipids.
Using of amino acids:
1) for protein synthesis;
2) for synthesis of nitrogen containing compounds (creatine, purines,
choline, pyrimidine);
3) as the source of energy (oxidation deamination, transamination,
decarboxilation);
4) for the gluconeogenesis;
5) for the formation of biologically active compounds.
Enzymes,which catalyse these
processes are called
transaminases or
aminotransferases. They require
pyrudocal phosphate (vit. B6) as
the coenzyme it acts as a carrier
of amino group.
AlAT is predominent in liver, AsAT
in heart.

General scheme of transamination
R
1
C NH
2
H
COOH
COOH
CH
2
CH
2
HC NH
2
COOH
COOH
CH
2
CH
2
C O
COOH
R
1
C O
COOH
-Amino acid
-ketoglutarate
-glutamate
-Keto acid
Amino-
transfe-
rase
Deamination of amino acids.
Deamination - eliminating of amino group from
amino acid with ammonia formation. There are
four types of deamination: oxidative, reduction,
hydrolitic and intramolecular.
Oxidative deamination. Some amino acids, for
example glutamate, may undergo oxidative
deamination catalyzed by the pyridine-linked
dehydrogenase, which is present in both the
cytosol and mitochondria of the liver.
The NADH formed is ultimately oxidized by the
electron-transport chain. L-Glutamate
dehydrogenase plays a central role in amino acid
deamination because in most organisms glutamate
is the only amino acid that has such an active
dehydrogenase.

OXIDATIVE DEAMINATION OF AMINO ACIDS
COOH
CH
2
CH
2
HC NH
2
COOH

+
+
+
(1)
COOH
(2)
CH
2
CH
2
C NH
COOH
H
2
O
COOH
CH
2
CH
2
C O
COOH
+ NH
3
-glutamate
-ketoglutarate -iminoglutarate
Glutamate
dehydroge
nase
Reduction deamination:
R-CH(NH2)-COOH + NADH2 R-CH2-COOH
+ NH3 + NAD+
Amino acid fatty acid
Hydrolitic deamination:
R-CH(NH2)-COOH + H2O R-CH(OH)-COOH
+ NH3
Amino acid oxiacid
Intramolecular deamination:
R-CH2-CH(NH2)-COOH R-CH-CH-COOH +
NH3
Amino acid unsaturated fatty acid
Indirect deamination of amino acids
-Amino acid
+
-ketoglutarate
-Keto acid
+
-glutamate
deamination
+ NAD
-NADH+H
-NH
3
Decarboxilation non reversible
process.
Amino acides are decarboxilated to
give amines. The enzyme is
decarboxilase, it requiers pyridoxal
phosphste as cofactor.
Bioligically active amines (biogenic
amines)
1. Histidine histamine
2. Glutamic acid - aminobutyric
acid (GABA)
3. Tryptophan serotonin
4. Tyrosine adrenalin, noradrenalin
1) A lot of histamine is formed in inflamatory place;
It has vasodilator action;
Mediator of inflamation, mediator of pain;
Responsible for the allergy development;
Stimulate HCI secretion in stomach.
-CO2
2) Tryptophan Serotonin
Vasokonstrictor
Takes part in regulation of arterial pressure, body
temperature, respiration, kidney filtration, mediator of
nervous system
3) Tyrosine Dopamine
It is precursor of epinephrine and norepinephrine.
mediator of central nervous system
4) Glutamate -aminobutyrate (GABA)
Is is ingibitory mediator of central nervous system. In
medicine we use with anticonvulsion purpose (action).
DECARBOXYLATION OF AMINO ACIDS
R CH COOH
NH
2
CO
2
R CH
2
NH
2
decarboxylase
amine
-Amino acid
COOH
HC NH
2
CH
2
CH
2
COOH
CO
2
NH
2
CH
2
CH
2
CH
2
COOH
-amino
butyrate
-glutamate
C
N
H
N CH
2
CH COOH
NH
2
CO
2
C
N
H
N
CH
2
CH
2
NH
2
Histamine
Histidine
The urea cycle
Urea formation, which takes place in the liver
of ureotelic organisms, is brought about by
the urea cycle, a cyclic pathway first
postulated by H.A.Krebs.
The first amino group entering the urea cycle
arises as free ammonia following the
oxidative deamination of glutamate in liver
mitochondria.
The free ammonia so formed is then utilized,
together with carbon dioxide, to form
carbamoyl phosphate, a very unstable
compound, in a complex reaction catalyzed
by carbamoyl phosphate synthetase
(ammonia), present in the mitochondrial
matrix:
SYNTHESIS OF UREA
NH
3
+ CO
2
+ 2 + H
2
O
H
2
N C
O
O ~ P
OH
OH
O + 2 + H
3
PO
4
Carbamoilphosphate synthase
carbamilphosphate
Two molecules of ATP are required to form
each molecule of carbamoyl phosphate in this
reaction, which is essentially irreversible. The
formation of carbamoyl phosphate by this
pathway in the mitochondria is specialized for
urea synthesis.
The carbamoyl phosphate generated in the
mitochondria now donates its carbamoyl
group to ornithine, which is formed in the
cytosol but enters the mitochondrion via a
specific inner-membrane transport system.
The product is citruiline:
H
2
N C
O
O ~ P
OH
OH
O
+
NH
2
(CH
2
)
3
HC NH
2
COOH
NH
(CH
2
)
3
HC NH
2
COOH
C O
NH
2
+ H
3
PO
4
SYNTHESIS OF UREA
carbamilphosphate
ornitine
citrulline
Ornitine-carbomoil transferasa
This reaction is catalyzed by ornithine
carbamoyltransferase of the mitochondrial
matrix. The citrulline formed now leaves the
mitochondrial matrix and passes to the
cytosol, where the remaining reactions of the
urea cycle take place.
The second amino group required for urea
synthesis now arrives in the form of
aspartate, which in turn acquired it from
glutamate by the action of aspartate
transaminase in the cytosol. The amino group
of aspartate condenses reversibly with the
carbamoyl carbon atom of citrulline in the
presence of ATP to form argininosuccinate;
this reaction is catalyzed by argminosuccinate
synthetase:
NH
(CH
2
)
3
HC NH
2
COOH
C NH
NH
CH
COOH
CH
2
COOH
NH
(CH
2
)
3
HC NH
2
COOH
C NH
NH
2
CH
COOH
CH
COOH
+
SYNTHESIS OF UREA
arginino-succinate
arginine
fumarate
Arginino-succinate lyase
In the next reaction argininosuccinate
undergoes a elimination reaction by
the action of argininosuccinate lyase to
form free arginine and fumarate:
SYNTHESIS OF UREA
NH
(CH
2
)
3
HC NH
2
COOH
C O
NH
2
COOH
CH
CH
2
H
2
N
COOH
+ +
NH
(CH
2
)
3
HC NH
2
COOH
C N
NH
2
CH
COOH
CH
2
COOH
NH
(CH
2
)
3
HC NH
2
COOH
C NH
NH
CH
COOH
CH
2
COOH
+ M + H
4
P
2
O
7
citrulline
aspartate
arginino-succinate
Arginino-succinate synthase
The arginine formed in this reaction
becomes the immediate precursor of
urea, whereas the fumarate returns to
the pool of tricarboxylic acid cycle
intermediates.
Up to this point the reaction sequence
is that employed by all organisms
capable of the biosynthesis of arginine.
However, only ureotelic animals
possess large amounts of arginase,
which cleaves urea from arginine and
regenerates ornithine, a reaction taking
place in the cytosol:
SYNTHESIS OF UREA
NH
(CH
2
)
3
HC NH
2
COOH
C NH
NH
2
+ H
2
O
NH
2
(CH
2
)
3
HC NH
2
COOH
+ C O
NH
2
NH
2
arginine
ornitine
urea
arginase
Diagnostic significance of the
determination of urea in urine.
25-30 g/day of urea is excreted in
normal conditions.
The increase of urea in urine
occurs in high fever, malignant
anemia, poisoning by phosphorus,
intensive decomposition of protein
in organism. The decrease of urea
in urine occurs in liver diseases,
kidney unsufficiency, acidosis.