Amino Acid 1

Chapter 17
Amino Acid Oxidation and the

Production of Urea
1
zhengzhou univercity 12/01/09
amino acids can undergo oxidative
degradation in three different metabolic
circumstances
 When a diet is rich in protein, and amino acids are ingested
in excess of the body's needs for protein synthesis, the
surplus may be catabolized; amino acids cannot be stored.
 During the normal synthesis and degradation of cellular
proteins (protein turnover) some of the amino acids
released during protein breakdown will undergo oxidative
degradation if they are not needed for new protein
synthesis.
 During starvation or in diabetes mellitus, when
carbohydrates are either unavailable or not properly
utilized, body proteins are called upon as fuel. 2
Digest
ion
3
degradation of ingested proteins in the
stomach.
Entry of protein into the stomach stimulates the gastric
mucosa to secrete the hormone gastrin, which in turn
stimulates the secretion of hydrochloric acid (pH 1.5 to
2.5 denature, antiseptic) and pepsinogen.
Pepsinogen :an inactive precursor or zymogen is
converted into active pepsin in the gastric juice by the
enzymatic action of pepsin.42 amino acid residues are
removed from the amino-terminal end of
Pepsinogen.pepsin hydrolyzes ingested proteins at
peptide bonds on the amino-terminal side of the
aromatic amino acid residues Tyr, Phe, and Trp
,cleaving long polypeptide chains into a mixture of
smaller peptides.
4
degradation of ingested proteins in the
small intestine.
 As the acidic stomach contents pass into the
small intestine, the low pH triggers the secretion
of the hormone secretin into the blood. Secretin
stimulates the pancreas to secrete bicarbonate
into the small intestine to neutralize the gastric
HCI, increasing the pH abruptly to about pH 7.
 The entry of smaller peptides into the upper
part of the intestine (duodenum) releases the
hormone cholecystokinin, which stimulates
secretion of several pancreatic enzymes. Three of
these, trypsin, chymotrypsin, and
carboxypeptidase, are made by pancreas as their
respective enzymatically inactive zymogens,
trypsinogen, chymotrypsinogen, and
procarboxypeptidase.
5
Zymogens actived
 After trypsinogen enters the small
intestine, it is converted into its active
form, trypsin. Once some free trypsin has
been formed, it also can catalyze the
conversion of trypsinogen into trypsin
.Trypsin, as noted above, can convert
chymotrypsinogen and
procarboxypeptidase into chymotrypsin
and carboxypeptidase.
6
Zymogen?
Synthesis of these enzymes as inactive
precursors protects the exocrine cells from
destructive proteolytic attack.
The pancreas protects itself against self
digestion in another way-by making a
specific inhibitor, itself a protein, called
pancreatic trypsin inhibitor (p. 235).
7
disease
 acute pancreatitis. In this condition,
caused by obstruction of the normal
pathway of secretion of pancreatic juice
into the intestine, the zymogens of the
proteolytic enzymes are converted into
their catalytically active forms
prematurely, inside the pancreatic cells. As
a result these powerful enzymes attack the
pancreatic tissue itself, causing a painful
and serious destruction of the organ, which
can be fatal.
8
Degradation of the short peptides
in the small intestine by other
peptidases
 carboxypeptidase, which removes
successive carboxyl-terminal residues
from peptides.
 aminopeptidase, which can hydrolyze
successive amino-terminal residues from
short peptides.
 Endopeptidase and exopeptidase
9
Amino acid +
10
Absorption
 By the sequential action of these proteolytic
enzymes and peptidases, ingested proteins
are hydrolyzed to yield a mixture of free
amino acids, which can then be transported
across the epithelial cells lining the small
intestine
 The free amino acids enter the blood
capillaries in the villi and are transported to
the liver.
11
Main use of amino acid
 Proteins are chains of amino acids, each
joined to its neighbor by a specific type of
covalent bond. (Protein synthesis )
12
nonessential amino acids and essential
amino acids :available for biosynthesis
13
Metabolic Fates of Amino acid
..
14
a-keto acids
 Under different circumstances, amino
acids lose their amino groups, and the a-
keto acids so formed may undergo
oxidation to CO2 and H2O. In addition,
and often equally important, the carbon
skeletons of the amino acids provide three-
and four-carbon units that can be
converted to glucose, which in turn can
fuel the functions of the brain, muscle, and
other tissues
15
Metabolic Fates of ammonia
 Most of the amino acids are metabolized in the
liver. Some of the ammonia that is generated is
recycled and used in a variety of biosynthetic
processes;
 the excess is either excreted directly(kidney) or
converted to urea for excretion, depending on the
organism. Excess ammonia generated in other
(extrahepatic) tissues is transported to the liver in
the form of amino groups, for conversion to the
appropriate excreted form(urea)
16
Glutamine Carries Ammonia to
the Liver
 Ammonia is quite toxic to animal tissues .In most
animals excess ammonia is converted into a
nontoxic compound before export from
extrahepatic tissues into the blood and thence to
the liver or kidneys.. Glutamine is a nontoxic,
neutral compound that can readily pass through
cell membranes.the amide nitrogen is released as
ammonia only within liver mitochondria, where
the enzyme glutaminase converts glutamine to
glutamate and NH4+ .
17
transport of amino groups
:glutamate and glutamine
 The amino acids glutamate and glutamine play
especially critical roles in these pathways .Amino
groups from amino acids are generally first
transferred to α-ketoglutarate in the cytosol of
liver cells to form glutamate. Glutamate is then
transported into the mitochondria;Excess
ammonia generated in most other tissues is
converted to the amide nitrogen of glutamine,
then transported to liver mitochondria. In most
tissues, one or both of these amino acids are
found in elevated concentrations relative to other
amino acids.
18
Glutamine is a major transport form of ammonia; it is normally present
in blood in much higher concentrations than other amino acids.
19
20
Alanine Carries Ammonia from
Muscles to the Liver
 The use of alanine to transport ammonia
from hard-working skeletal muscles to the
liver. Vigorously contracting skeletal
muscles operate anaerobically, producing
not only ammonia from protein breakdown
but also large amounts of pyruvate from
glycolysis. Both these products must find
their way to the liver.ammonia to be
converted into urea for excretion and
pyruvate to be rebuilt into glucose.
21
The glucose-
alanine
cycle:Alanine serves
as a carrier of ammonia
and of the carbon skeleton
of pyruvate from muscle to
liver. The ammonia is
excreted, and the pyruvate
is used to produce
glucose, which is returned
to the muscle.
Gluconeogen
esis
22
glucose-alanine cycle
 Animals solve two problems with one
cycle: they move the carbon atoms of
pyruvate, as well as excess ammonia, from
muscle to liver as alanine. In the liver,
alanine yields pyruvate, the starting
material for gluconeogenesis, and releases
NH4+ for urea synthesis. Alanine also
plays a special role in transporting amino
groups to the liver in a nontoxic form, by
the glucose-alanine cycle
23
Overview of amino group catabolism in the liver Excess
NH4+ is excreted as urea or uric acid.
24
The aminotransferase reaction (transamination). α-ketoglutarate is the
amino group acceptor. All aminotransferases have pyridoxal phosphate
(PLP) as cofactor.
25
Pyridoxal phosphate (PLP, vitamin B6) and its
aminated form pyridoxamine phosphate are the
tightly bound coenzymes of aminotransferases.
26
transamination
 Cells contain several different aminotransferases,
many specific for a-ketoglutarate as the amino
group acceptor. The aminotransferases differ in
their specificity for the other substrate, the L-
amino acid that donates the amino group, and are
named for the amino group donor .The reactions
catalyzed by the aminotransferases are freely
reversible, having an equilibrium constant of
about 1.0 .
27
Aminotransferases
 Aminotransferases are classic examples of
enzymes catalyzing bimolecular ping-pong
reactions .In such reactions the first substrate
must leave the active site before the second
substrate can bind. Thus the incoming amino
acid binds to the active site, donates its amino
group to pyridoxal phosphate, and departs in the
form of an a-keto acid. Then the incoming a-keto
acid is bound, accepts the amino group from
pyridoxamine phosphate, and departs in the form
of an amino acid.
28
Aminotransferase is a enzyme in
cell
 The measurement of alanine
aminotransferase and aspartate
aminotransferase levels in blood serum is
an important diagnostic procedure in
medicine, used as an indicator of heart
damage and to monitor recovery from the
damage
29
diagnosis
 Alanine aminotransferase (ALT; also called glutamate-
pyruvate transaminase, GPT) and glutamate-oxaloacetate
transaminase, GOT) are important in the diagnosis of heart
and liver damage. Occlusion of a coronary artery by lipid
deposits can cause severe local oxygen starvation and
ultimately the degeneration of a localized portion of the
heart muscle; Such damage causes aminotransferases,
among other enzymes, to leak from the injured heart cells
into the bloodstream. Measurements of the concentration
in the blood serum of these two aminotransferases by the
SGPT and SGOT tests (S for serum) can provide
information about the severity and the stage of the damage
to the heart.
30
 SGOT and SGP'f are also important in industrial
medicine to determine whether people exposed
to carbon tetrachloride, chloroform, or other
solvents used in the chemical, dry-cleaning, and
other industries have suffered liver damage.
These solvents cause liver degeneration, with
resulting leakage into the blood of various
enzymes from the injured hepatocytes.
Aminotransferases, because they are very active
in liver and their activity can be detected in very
small amounts, are most useful in the monitoring
of people exposed to such industrial chemicals
31
The essential amino acids (those with carbon
skeletons that cannot be synthesized by animals
and must be obtained in the diet)
32
oxidative deamination
Glutamate is transported from the cytosol to the mitochondria, where it undergoes oxidative
deamination catalyzed by L-glutamate dehydrogenase . This enzyme, which is present
only in the mitochondrial matrix, requires NAD+ (or NADP+) as the acceptor of the
reducing equivalents .
33
transdeamination
 The combined action of the aminotransferases and
glutamate dehydrogenase is referred to as
transdeamination.
34
Ammonia Is Toxic to Animals
 The catabolic production of ammonia

poses a serious biochemical problem
because ammonia is very toxic. The
terminal stages of ammonia intoxication in
humans are characterized by the onset of a
comatose state and other effects on the
brain, The major toxic effects of ammonia
in brain probably involve changes in
cellular pH and the depletion of certain
citric acid cycle intermediates.
35
 The protonated form of ammonia
(ammonium ion) is a weak acid, and the
unprotonated form is a strong base:Most of
the ammonia generated in catabolism is
present as NH4+ at neutral pH. Although
many of the reactions that produce
ammonia, such as the glutamate
dehydrogenase reaction, yield NH4+ .
36
 Depletion of glutamate in the glutamine
synthetase reaction may have additional effects
on the brain. Glutamate, and the compound γ-
aminobutyrate (GABA) that is derived from it ,
are both important neurotransmitters; the
sensitivity of the brain to ammonia may well
reflect a depletion of neurotransmitters as well
as changes in cellular pH and ATP metabolism.
37
38
Summary
A small fraction of oxidative energy
in humans comes from the catabolism
of amino acids. Amino acids are
derived from the normal breakdown
(recycling) of cellular proteins,
degradation of ingested proteins, or
breakdown of body proteins in lieu of
other fuel sources during starvation or
in untreated diabetes mellitus.
39
 Ingested proteins are degraded in the
stomach and small intestine by proteases.
Most proteases are initially synthesized as
inactive zymogens, which are activated in
the stomach or intestine by proteolytic
removal of parts of their polypeptide
chains.
40
 An early step in the catabolism of amino acids is
the separation of the amino group from the
carbon skeleton.
 In most cases, the amino group is transferred to
α-ketoglutarate to form glutamate. This type of
reaction is called a transamination and requires
the coenzyme pyridoxal phosphate.
 Glutamate is transported to liver mitochondria,
where an amino group is liberated as ammonia
(NH4+ ) by the enzyme glutamate
dehydrogenase.
41
Ammonia formed in other tissues is
transported to liver mitochondria as the
amide nitrogen of glutamine or as the
amino group of alanine. Most of the
alanine is generated in muscle and
transported in the blood to the liver. After
deamination the resulting pyruvate is
converted to glucose, which is
transported back to muscle as part of the
glucose-alanine cycle.
42
problems ( Answer )
 1. Products of Amino Acid

Transamination Draw the structure and
give the name of the α-keto acid resulting
when the following amino acids undergo
transamination with α-ketoglutarate:
 (a) Aspartate
(c) Alanine
(b) Glutamate
(d) Phenylalanine
43
 2. Measurement of the Alanine
Aminotransferase Reaction Rate The
activity (reaction rate) of alanine
aminotransferase is usually measured by
including an excess of pure lactate
dehydrogenase and NADH in the reaction
system. The rate of alanine disapperance is
equal to the rate of NADH disappearance
measured spectrophotometrically. Explain
how this assay works.
44
 3. Distribution of Amino Nitrogen If your
diet is rich in alanine but deficient in
aspartate, will you show signs of aspartate
deficiency? Explain.
45
 4. A Genetic Defect in Amino Acid Metabolism: A Case History A
two-year-old child was brought to the hospital. His mother indicated
that he vomited frequently, especially after feedings. The child's
weight and physical development were below normal. His hair,
although dark, contained patches ot white. A urine sample treated
with ferric chloride (FeCl3) gave a green color characteristic of the
presence of phenylpyruvate. Quantitative analysis of urine samples
gave the results shown in the table below.
 (a) Suggest which enzyme might be deficient. Propose a treatment
for this condition.
(b) Why does phenylalanine appear in the urine in large amounts?
(c) What is the source of phenylpyruvate and phenyllactate? Why
does this pathway (normally not functional) come into play when the
concentration of phenylalanine rises?
(d) Why does the patient's hair contain patches of white?
46

Amino Acid 1

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Chapter 17

Amino Acid Oxidation and the

NH4+ is excreted as urea or uric acid.

 The catabolic production of ammonia

 1. Products of Amino Acid

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