Chapter 18
Amino Acid Oxidation and
the Production of Urea
1,The surplus amino acids in
animals can be completely oxidized
or converted to other storable fuels
? Amino acids in excess (from diet,protein turnover)
can neither be stored,nor excreted,but oxidized to
release energy or converted to fatty acids or glucose.
? Animals also utilize amino acid for energy
generation during starvation or in diabetes mellitus.
? Microorganisms can also use amino acids as an
energy source when the supply is in excess.
? Plants almost never use amino acids as an energy
source (neither fatty acids).
2,Dietary proteins are digested into
amino acids in the gastrointestinal
tract
? Pepsin cleaves polypeptides into smaller peptides in
stomach (N-terminal side of Y,F,W residues).
? Trypsin (C-terminal side of K,R) and chymotrypsin
(C-terminal side of F,W,and Y) further cleave the
peptides in small intestine.
? Carboxypeptidase and aminopeptidase cleave the
small peptides into amino acids,which are then
absorbed and eventually delivered to liver.
Pepsin
Chymotrypsin,trypsin,
and exopeptidases
Amino acids
3,The amino groups and carbon
skeletons of amino acids take separate
but interconnected pathways
? The amino group is reused or excreted,as ammonia,
urea (via the urea cycle) or uric acid.
? The carbon skeletons (a-keto acids) generally find
their way to the citric acid cycle for further
oxidation or conversion.
? The degradation of the carbon skeletons may be
very complicated but similar to that of fatty acids in
some cases.
4,Liver is the major site of amino
acid degradation in vertebrates
? a-ketoglutarate (from amino acids),glutamate (from
free ammonia),pyruvate (from muscle amino acids)
collects amino groups (in forms of Glu,Gln and Ala)
to liver mitochondria for further processing.
? The excess NH4+ is excreted directly in bony fishes,
as urea in most terrestrial vertebrates,or as uric acid
in birds and terrestrial reptiles.
? The carbons in both urea and uric acid are highly
oxidized (with most of the energy extracted).
5,PLP facilitates the transaminatin and
other transformations of amino acids
? Different aminotransferases (e.g.,aspartate and
alanine aminotransferases),each catalyzes the
transfer of the amino group from an amino acid to
a-ketoglutarate to form Glu and a a-keto acid.
? Pyridoxal phosphate (PLP),being derived from
vitamin B6 (i.e.,pyridoxine) and the prosthetic
groups for all the aminotransferases,act as a
temporary carrier of the amino groups.
? PLP accepts and then donates an amino group by
forming a Schiff base with the amino-donating
amino acid and amino-accepting a-keto acid (being
a-ketoglutarate in many cases )respectively.
? PLP is seen bound covalently to the active sites of
aminotransferases via structure determination.
? The reaction occurs via a typical ping-pong
mechanism,the first product leaves before the
second substrate enters.
? An interconversion between an aldimine（ 醛亚胺 ）
and a ketimine（ 酮亚胺 ） is proposed to occur via
a quinonoid（ 醌型的 ） intermediate during the
transamination reaction.
? PLP act in a variety of other reactions at the a,b,
and g carbons of amino acids,including
racemizations,decarboxylations at the a carbon,
elimination and replacement reactions at the b and g
carbons.
Aspartate aminotransferase (dimeric)
PLP
2-methyl-Asp-PLP
PLP accepts and donates
amino groups in the active
site of aminotransferases
Ketimine
PLP facilitates
transamination,
racemization,
decarboxylation
of amino acids
6,Glutamate collects and delivers
free ammonia to the liver
? Free ammonia is produced in tissues (e.g.,by the
direct deamination of Ser and Thr,deamination of
nucleotides) and is toxic to animals,thus need to be
transported to the liver by first converting to a
nontoxic compound,Gln.
? Glutamine synthetase catalyzes the two-step
addition of ammonia to Glu to yield Gln,with ATP
consumed to activate the g-COOH.
? This reaction also represents how fixed nitrogen in
bacteria enter biomolecules; Gln is the source of
amino groups in a variety of biosynthetic reactions.
7,The pyruvates and amino groups are
transported to liver in the form of Ala
? Pyruvate is an abundant product from muscle
glycolysis and can take an amino group (from Glu)
to form Ala (alternatively,be reduced to lactate).
? Alanine once gets to liver will then transfers it’s
amino group to a-ketoglutarate or oxaloacetate to
reform pyruvate.
? Pyruvate is reconverted to glucose in liver via the
gluconeogenesis pathway,which will be transported
back to muscle for energy supply.
? This is called alanine-glucose cycle,complementing
the Cori cycle for liver to continuously provide
glucose to vigorously contracting skeletal muscles.
Ala
Ala
Ala
Pyruvate
Pyruvate
The alanine-
glucose cycle
8,Gln and Glu releases NH4+ in
liver mitochondria
? The glutaminase in liver mitochondria catalyzes the
conversion of Gln to Glu,releasing NH4+ (from the
side chain amide of Gln).
? The glutamate dehydrogenase (a hexameric
allosteric enzyme) there catalyzes the oxidative
deamination of Glu,releasing NH4+,with released
electrons collected by either NAD+ or NADP+.
? Glutamate dehydrogenase is allosterically activated
by ADP and GDP,but inhibited by ATP and GTP,
i.e.,a lowering of the energy charge accelerates the
oxidation of amino acids.
9,NH4+ in hepatocytes is converted
into urea in most terrestrial
vertebrates for excretion
? Hans Krebs and Kurt Henseleith revealed in 1932
(five years before the citric acid cycle was
elucidated) that this conversion is accomplished via
a cyclic pathway,called the urea cycle.
? The two nitrogen atoms are derived from Asp and
NH4+,with the carbon atom coming from CO2.
? Ornithine（ 鸟氨酸 ）,acting as the carrier of the
carbon and nitrogen atoms,has a role similar to that
of oxaloacetate in citric acid cycle.
? The urea cycle spans the mitochondria and cytosol.
? Four phosphoanhydride bonds （ equivalent to four
ATP molecules) are hydrolyzed for the production
of each urea.
The urea cycle takes place in the mitochondria
and cytosol of liver cells
10,The conversion of ammonia to
urea takes five (six) enzymatic steps
? First a carbamoyl phosphate (氨基甲酰磷酸, an
activated carbamoyl group donor) is produced in the
mitochondrial matrix using one NH4+,one HCO3-,
and two ATP molecules,with the catalysis of
carbamoyl phosphate synthetase I (isozyme II exists
in the cytosol,acting in pyrimidine synthesis).
? The urea cycle begins in the mitochondrial matrix
when a carbamoyl group is transferred to ornithine
to form citrulline（ 瓜氨酸 ）,in a reaction
catalyzed by ornithine transcarbamoylase.
? Citrulline moves into the cytosol.
? The amino group of one Asp condenses with the
ureido group of citrulline to form argininosuccinate,
in an ATP-dependent reaction catalyzed by
argininosuccinate synthetase,via a citrullyl-AMP
intermediate.
? The argininosuccinate is then cleaved to form Arg
and fumarate,in a reaction catalyzed by
argininosuccinate lyase.
? Arg is then cleaved to form urea and regenerate
ornithine at the same time,catalyzed by arginase.
? Fumarate enters mitochondria and be reconverted to
Asp,be converted to oxaloacetate first via the citric
acid cycle (generating one NADH),then take an
amino group from Glu via a transamination reaction.
The synthesis of
carbamoyl phosphate
requires two activation
steps,consuming two
ATP molecules
The urea cycle and citric
acid cycle are linked via
the aspartate-arginino-
succinate shunt
11,The rate of urea synthesis is
controlled at two levels
? Allosteric regulation of enzyme activity (fast),N-
acetylglutamate,synthesized from acetyl-CoA and
glutamate by the catalysis of N-acetylglutamate
synthase,positively regulates carbamoyl phosphate
synthetase I activity.
? Gene regulation of enzyme amount (slow),
syntheses of the five liver enzymes involved in urea
synthesis are increased during starvation (when
energy has to be obtained from muscle proteins!) or
after high protein uptake.
? The rates of transcription of the five genes encoding
the enzymes are increased.
Carbamoyl phosphate
synthetase I is positively
regulated by the enigmatic
N-acetyl-Glutamate
12,Genetic defects of the urea
production enzymes lead to
hyperammonemia and brain damage
? Genetic defects of all five urea cycle enzymes have
been revealed.
? High levels of ammonia lead to mental disorder or
even coma and death.
? Ingenious strategies for coping with the deficiencies
have been devised based on a thorough
understanding of the underlying biochemistry.
? Strategy I,diet control,provide the essential amino
acids in their a-keto acid forms.
? Strategy II,when argininosuccinate lyase is
deficient,ingesting a surplus of Arg will help
(ammonia will be carried out of the body in the form
of argininosuccinate,instead of urea).
? Strategy III,when carbamoyl phosphate synthetase,
ornithine transcarbamoylase,or argininosuccinate
sythetase are deficient,the ammonia can be
eliminated by ingesting compounds (e.g.,benzoate
or phenylacetate),which will be excreted after
accepting ammonia.
13,The carbon skeletons of the amino
acids are first converted (funneled) into
seven major metabolic intermediates
? These are acetoacetyl-CoA,acetyl-CoA (which can
be converted to fatty acids and ketone bodies,the
amino acids are thus ketogenic); pyruvate,a-
ketoglutarate,succinyl-CoA,fumarate,oxaloacetate
(all can be converted to glucose,the amino acids are
thus glucogenic).
? Amino acids having long carbon chains often
degrade in ways similar to fatty acid oxidation.
? Amino acids having short chains often degrade into
pyruvate or citric acid intermediates directly.
? Some of the conversions are very complicated,
using a variety of cofactors.
The carbon skeletons of
amino acids are oxidized via
the citric acid cycle
14,Tetrahydrofolate (H4 folate) acts
as a carrier for activated one-carbon
units in amino acid metabolism
? The cofactor,derived from folate (a vitamin),
consists of three moiety,a 6-methylpterin,a p-
aminobenzoate,and a glutamate.
? It usually carries one-carbon unit of intermediate
oxidation levels,methylene; methenyl,formyl
formimino group,and sometimes methyl groups.
? The most reduced and oxidized one carbon forms,
methyl group and CO2 are usually carried by S-
adenosylmethionine and biotin respectively.
? The one-carbon unit is attached to N-5,or/and N-10
on tetrahydrofolate,
? The different forms of the attached one-carbon units
are interconvertible.
? Tetrahydrofolate works in amino acid catabolism
(e.g.,Gly,His) and anabolism (e.g.,Ser,Met,Gly).
? It is also used in purine and pyrimidine syntheses.
Specific carriers carry and transfer various
one-carbon units
Tetrahydrofolate
carries and
interconverts
one carbon
units of
intermediate
oxidation state
15,Some amino acids are converted to
intermediates of citric acid cycle by
simple removal of the amino groups
? Ala to pyruvate; Asp to oxaloacetate; Glu to a-
ketoglutarate; Asp to oxaloacetate; Glu to a-
ketoglutarate.
? Asn is converted to Asp via a deamination reaction
catalyzed by asparaginase.
? Gln is converted to Glu also via a deamination
reaction catalyzed by glutaminase,
16,Gly can be converted to Ser or
CO2 and NH4+ for oxidation in
different organisms?
In animals,Gly is oxidized to form CO2 and NH4+,
with electrons collected by NAD+ and one carbon
unit transferred to H4 folate (to form N5,N10-
methylene-H4 folate) in a reaction catalyzed by
glycine synthase.
? In microorganisms,Gly is first converted to Ser by
accepting a hydroxymethyl group from N5,N10-
methylene-H4 folate in a reaction catalyzed by serine
hydroxymethyl transferase; Ser is then converted to
pyruvate for further oxidation.
Predominant
in animalsPredominant in bacteria
Two metabolic fates of glycine
17,Acetyl-CoA is formed from the
degradation of many amino acids
? Trp,Ala,Ser,Gly and Cys,are converted to
pyruvate first before being converted to acetyl-CoA.
? Portions of the carbon skeletons of Trp,Phe,Try,
Lys,Leu,and Ile are converted (via acetoacetyl-CoA
or directly)to acetyl-CoA.
? Some of the final steps in the degradation of Leu,
Lys,and Trp resemble steps in fatty acid oxidation.
? Tryptophan degradation is the most complex.
? Intermediates of tryptophan degradation are
precursors for synthesizing other biomolecules,
including nicotinate,serotonin,and indoleacetate.
Trp,Ala,Ser,Gly and Cys are converted to
pyruvate,before being converted to acetyl-CoA
Trp,Phe,Try,
Lys,Leu,and Ile
are also converted
to acetyl-CoA
Trp is an important
biosynthetic
precursor
18,O2 is used to break the aromatic
rings of Phe and Tyr
? Phe is converted to Tyr by hydroxylation,in a
reaction catalyzed by phenylalanine hydroxylase,a
mixed-function oxygenase (or monooxygenase)
using tetrahydrobiopterin (a pterin derivative) as the
reductant.
? Mixed-function oxygenase catalyzes the
incorporation of the two oxygen atoms of O2 into
two different products (usually with one being H2O).
? Participating the conversion are also two
dioxygenases,which catalyze the incorporation of
both oxygen atoms of O2 into one product.
? Phe and Tyr are eventually converted to
fumarate and acetoacetyl-CoA.
Action of a dioxygenase,p-hydroxyphenyl-
pyruvate is converted to homogentisate (尿黑酸 ）
via a peracid and an epoxide intermediate; both
atoms of O2 emerge in homogentisate;
19,A few genetic diseases are related to
defects of Phe catabolism enzymes
? Defects in enzymes of amino acid degradation lead
to accumulation of specific intermediates,many of
which cause diseases.
? Deficiency of phenylalanine hydroxylase causes
phenylketonuria (PKU,苯丙酮尿症 ),one of the
earliest human genetic defects of metabolism (first
revealed in 1934 in testing mentally retarded
patients,whose urine turned olive-green on addition
of FeCl3),
? The accumulated Phe is converted to
phenylpyruvate (via an aminotransferase) which
caused the color change or urine.
? Another disease related to Phe/Tyr catabolism and
of historical interest is alkaptonuria (尿黑酸症 ).
? Garrod discovered that the condition is transmitted
as a single recessive inheritable trait and could be
traced to the absence of a single enzyme (early
1900s),thus hinted the direct relation between genes
and enzymes
? He recognized that homogentisate (尿黑酸 ),the
compound that is excreted in large amount and
whose oxidation turns the urine black,is a normal
intermediate in the degradation of Phe and Tyr;
? Homogentisate dioxygenase was found to be
defective in such patients.
Phenylalanine that is
accumulated in PKU
patients is
converted to other
alternative products.
20,Leu,Ile,and Val are degraded via
reactions similar to fatty acid oxidation
? The same branched-chain aminotransferase and
branched-chain a-keto acid dehydrogenase complex
catalyzes the first two degradative reactions
(transamination and oxidative decarboxylation) of
all these three amino acids in extrahepatic tissues.
? The a-keto acid dehydrogenase complex has a
similar structure and catalyzes essentially the same
type of reaction (oxidative decarboxylation of a-
keto acids) as pyruvate dehydrogenase complex and
the a-ketoglutarate dehydrogenase complex.
? The subsequent degradation of the a-keto acids
share similar strategies of fatty acid oxidation.
? Leu is finally converted to acetyl-CoA and
acetoacetate; Val to propionyl-CoA; Ile to acetyl-
CoA and propionyl-CoA.
? The propionyl-CoA produced is converted to
succinyl-CoA via carboxylation and intramolecular
rearrangement involving free radicals using
coenzyme B12 (as previously described).
? Defect of branched-chain a-keto acid
dehydrogenase complex leads to maple syrup urine
disease,all three a-keto acids and amino acids are
accumulated and the urine of the patients smell like
maple syrup (槭糖浆 ).
Summary
? Amino acid in excess can neither be stored,nor
excreted,but oxidized or converted.
? The amino groups and carbon skeletons of amino
acids take separate but interconnected pathways.
? Liver is the major site of amino acid degradation in
vertebrates.
? PLP facilitates the transaminatin and other
transformations of amino acids.
? Glutamate collects and delivers free ammonia to the
liver.
? Gln and Glu releases NH4+ in liver mitochondria.
? NH4+ in hepatocytes is converted into urea through
the urea cycle in most terrestrial vertebrates for
excretion.
? The conversion of ammonia to urea takes five (six)
enzymatic steps.
? The rate of urea synthesis is controlled at two levels.
? The carbon skeletons of the amino acids are first
converted (funneled) into seven major metabolic
intermediates.
? Some amino acids are converted to intermediates of
citric acid cycle by simple removal of the amino
groups.
? Acetyl-CoA is formed from the degradation of many
amino acids.
? O2 is used to break the aromatic rings of Phe and
Tyr.
? A few genetic diseases are related to defects of Phe
catabolism enzymes.
? Leu,Ile,and Val are degraded via reactions similar
to fatty acid oxidation
References
? Fitzpatrick,P,F,(1999),Tetrahydropterin-
dependent amino acid hydroxylases” Annu,Rev,
Biochem,68:355-382.
? Kirsch,J,F…,And Christen,P,(1984),Mechanism
of action of aspartate aminotransferase proposed on
the basis of its spatial structure” J,Mol,Biol,
174:497-525.
? Trchinsky,Y,M,(1989),Transamination,Its
discovery,biological and chemical aspects” Trends
Biochem,Sci,12:115-117.
? Halpern,J,(1985),Mechanisms of coenzyme B12-
dependent rearrangements” Science 227:869-875.
? McPhalen,C,A….,and Chothia,C,(1992),Domain
closure in mitochondrial aspartate aminotransferases”
J,Mol,Biol,227:197-213.
Amino Acid Oxidation and
the Production of Urea
1,The surplus amino acids in
animals can be completely oxidized
or converted to other storable fuels
? Amino acids in excess (from diet,protein turnover)
can neither be stored,nor excreted,but oxidized to
release energy or converted to fatty acids or glucose.
? Animals also utilize amino acid for energy
generation during starvation or in diabetes mellitus.
? Microorganisms can also use amino acids as an
energy source when the supply is in excess.
? Plants almost never use amino acids as an energy
source (neither fatty acids).
2,Dietary proteins are digested into
amino acids in the gastrointestinal
tract
? Pepsin cleaves polypeptides into smaller peptides in
stomach (N-terminal side of Y,F,W residues).
? Trypsin (C-terminal side of K,R) and chymotrypsin
(C-terminal side of F,W,and Y) further cleave the
peptides in small intestine.
? Carboxypeptidase and aminopeptidase cleave the
small peptides into amino acids,which are then
absorbed and eventually delivered to liver.
Pepsin
Chymotrypsin,trypsin,
and exopeptidases
Amino acids
3,The amino groups and carbon
skeletons of amino acids take separate
but interconnected pathways
? The amino group is reused or excreted,as ammonia,
urea (via the urea cycle) or uric acid.
? The carbon skeletons (a-keto acids) generally find
their way to the citric acid cycle for further
oxidation or conversion.
? The degradation of the carbon skeletons may be
very complicated but similar to that of fatty acids in
some cases.
4,Liver is the major site of amino
acid degradation in vertebrates
? a-ketoglutarate (from amino acids),glutamate (from
free ammonia),pyruvate (from muscle amino acids)
collects amino groups (in forms of Glu,Gln and Ala)
to liver mitochondria for further processing.
? The excess NH4+ is excreted directly in bony fishes,
as urea in most terrestrial vertebrates,or as uric acid
in birds and terrestrial reptiles.
? The carbons in both urea and uric acid are highly
oxidized (with most of the energy extracted).
5,PLP facilitates the transaminatin and
other transformations of amino acids
? Different aminotransferases (e.g.,aspartate and
alanine aminotransferases),each catalyzes the
transfer of the amino group from an amino acid to
a-ketoglutarate to form Glu and a a-keto acid.
? Pyridoxal phosphate (PLP),being derived from
vitamin B6 (i.e.,pyridoxine) and the prosthetic
groups for all the aminotransferases,act as a
temporary carrier of the amino groups.
? PLP accepts and then donates an amino group by
forming a Schiff base with the amino-donating
amino acid and amino-accepting a-keto acid (being
a-ketoglutarate in many cases )respectively.
? PLP is seen bound covalently to the active sites of
aminotransferases via structure determination.
? The reaction occurs via a typical ping-pong
mechanism,the first product leaves before the
second substrate enters.
? An interconversion between an aldimine（ 醛亚胺 ）
and a ketimine（ 酮亚胺 ） is proposed to occur via
a quinonoid（ 醌型的 ） intermediate during the
transamination reaction.
? PLP act in a variety of other reactions at the a,b,
and g carbons of amino acids,including
racemizations,decarboxylations at the a carbon,
elimination and replacement reactions at the b and g
carbons.
Aspartate aminotransferase (dimeric)
PLP
2-methyl-Asp-PLP
PLP accepts and donates
amino groups in the active
site of aminotransferases
Ketimine
PLP facilitates
transamination,
racemization,
decarboxylation
of amino acids
6,Glutamate collects and delivers
free ammonia to the liver
? Free ammonia is produced in tissues (e.g.,by the
direct deamination of Ser and Thr,deamination of
nucleotides) and is toxic to animals,thus need to be
transported to the liver by first converting to a
nontoxic compound,Gln.
? Glutamine synthetase catalyzes the two-step
addition of ammonia to Glu to yield Gln,with ATP
consumed to activate the g-COOH.
? This reaction also represents how fixed nitrogen in
bacteria enter biomolecules; Gln is the source of
amino groups in a variety of biosynthetic reactions.
7,The pyruvates and amino groups are
transported to liver in the form of Ala
? Pyruvate is an abundant product from muscle
glycolysis and can take an amino group (from Glu)
to form Ala (alternatively,be reduced to lactate).
? Alanine once gets to liver will then transfers it’s
amino group to a-ketoglutarate or oxaloacetate to
reform pyruvate.
? Pyruvate is reconverted to glucose in liver via the
gluconeogenesis pathway,which will be transported
back to muscle for energy supply.
? This is called alanine-glucose cycle,complementing
the Cori cycle for liver to continuously provide
glucose to vigorously contracting skeletal muscles.
Ala
Ala
Ala
Pyruvate
Pyruvate
The alanine-
glucose cycle
8,Gln and Glu releases NH4+ in
liver mitochondria
? The glutaminase in liver mitochondria catalyzes the
conversion of Gln to Glu,releasing NH4+ (from the
side chain amide of Gln).
? The glutamate dehydrogenase (a hexameric
allosteric enzyme) there catalyzes the oxidative
deamination of Glu,releasing NH4+,with released
electrons collected by either NAD+ or NADP+.
? Glutamate dehydrogenase is allosterically activated
by ADP and GDP,but inhibited by ATP and GTP,
i.e.,a lowering of the energy charge accelerates the
oxidation of amino acids.
9,NH4+ in hepatocytes is converted
into urea in most terrestrial
vertebrates for excretion
? Hans Krebs and Kurt Henseleith revealed in 1932
(five years before the citric acid cycle was
elucidated) that this conversion is accomplished via
a cyclic pathway,called the urea cycle.
? The two nitrogen atoms are derived from Asp and
NH4+,with the carbon atom coming from CO2.
? Ornithine（ 鸟氨酸 ）,acting as the carrier of the
carbon and nitrogen atoms,has a role similar to that
of oxaloacetate in citric acid cycle.
? The urea cycle spans the mitochondria and cytosol.
? Four phosphoanhydride bonds （ equivalent to four
ATP molecules) are hydrolyzed for the production
of each urea.
The urea cycle takes place in the mitochondria
and cytosol of liver cells
10,The conversion of ammonia to
urea takes five (six) enzymatic steps
? First a carbamoyl phosphate (氨基甲酰磷酸, an
activated carbamoyl group donor) is produced in the
mitochondrial matrix using one NH4+,one HCO3-,
and two ATP molecules,with the catalysis of
carbamoyl phosphate synthetase I (isozyme II exists
in the cytosol,acting in pyrimidine synthesis).
? The urea cycle begins in the mitochondrial matrix
when a carbamoyl group is transferred to ornithine
to form citrulline（ 瓜氨酸 ）,in a reaction
catalyzed by ornithine transcarbamoylase.
? Citrulline moves into the cytosol.
? The amino group of one Asp condenses with the
ureido group of citrulline to form argininosuccinate,
in an ATP-dependent reaction catalyzed by
argininosuccinate synthetase,via a citrullyl-AMP
intermediate.
? The argininosuccinate is then cleaved to form Arg
and fumarate,in a reaction catalyzed by
argininosuccinate lyase.
? Arg is then cleaved to form urea and regenerate
ornithine at the same time,catalyzed by arginase.
? Fumarate enters mitochondria and be reconverted to
Asp,be converted to oxaloacetate first via the citric
acid cycle (generating one NADH),then take an
amino group from Glu via a transamination reaction.
The synthesis of
carbamoyl phosphate
requires two activation
steps,consuming two
ATP molecules
The urea cycle and citric
acid cycle are linked via
the aspartate-arginino-
succinate shunt
11,The rate of urea synthesis is
controlled at two levels
? Allosteric regulation of enzyme activity (fast),N-
acetylglutamate,synthesized from acetyl-CoA and
glutamate by the catalysis of N-acetylglutamate
synthase,positively regulates carbamoyl phosphate
synthetase I activity.
? Gene regulation of enzyme amount (slow),
syntheses of the five liver enzymes involved in urea
synthesis are increased during starvation (when
energy has to be obtained from muscle proteins!) or
after high protein uptake.
? The rates of transcription of the five genes encoding
the enzymes are increased.
Carbamoyl phosphate
synthetase I is positively
regulated by the enigmatic
N-acetyl-Glutamate
12,Genetic defects of the urea
production enzymes lead to
hyperammonemia and brain damage
? Genetic defects of all five urea cycle enzymes have
been revealed.
? High levels of ammonia lead to mental disorder or
even coma and death.
? Ingenious strategies for coping with the deficiencies
have been devised based on a thorough
understanding of the underlying biochemistry.
? Strategy I,diet control,provide the essential amino
acids in their a-keto acid forms.
? Strategy II,when argininosuccinate lyase is
deficient,ingesting a surplus of Arg will help
(ammonia will be carried out of the body in the form
of argininosuccinate,instead of urea).
? Strategy III,when carbamoyl phosphate synthetase,
ornithine transcarbamoylase,or argininosuccinate
sythetase are deficient,the ammonia can be
eliminated by ingesting compounds (e.g.,benzoate
or phenylacetate),which will be excreted after
accepting ammonia.
13,The carbon skeletons of the amino
acids are first converted (funneled) into
seven major metabolic intermediates
? These are acetoacetyl-CoA,acetyl-CoA (which can
be converted to fatty acids and ketone bodies,the
amino acids are thus ketogenic); pyruvate,a-
ketoglutarate,succinyl-CoA,fumarate,oxaloacetate
(all can be converted to glucose,the amino acids are
thus glucogenic).
? Amino acids having long carbon chains often
degrade in ways similar to fatty acid oxidation.
? Amino acids having short chains often degrade into
pyruvate or citric acid intermediates directly.
? Some of the conversions are very complicated,
using a variety of cofactors.
The carbon skeletons of
amino acids are oxidized via
the citric acid cycle
14,Tetrahydrofolate (H4 folate) acts
as a carrier for activated one-carbon
units in amino acid metabolism
? The cofactor,derived from folate (a vitamin),
consists of three moiety,a 6-methylpterin,a p-
aminobenzoate,and a glutamate.
? It usually carries one-carbon unit of intermediate
oxidation levels,methylene; methenyl,formyl
formimino group,and sometimes methyl groups.
? The most reduced and oxidized one carbon forms,
methyl group and CO2 are usually carried by S-
adenosylmethionine and biotin respectively.
? The one-carbon unit is attached to N-5,or/and N-10
on tetrahydrofolate,
? The different forms of the attached one-carbon units
are interconvertible.
? Tetrahydrofolate works in amino acid catabolism
(e.g.,Gly,His) and anabolism (e.g.,Ser,Met,Gly).
? It is also used in purine and pyrimidine syntheses.
Specific carriers carry and transfer various
one-carbon units
Tetrahydrofolate
carries and
interconverts
one carbon
units of
intermediate
oxidation state
15,Some amino acids are converted to
intermediates of citric acid cycle by
simple removal of the amino groups
? Ala to pyruvate; Asp to oxaloacetate; Glu to a-
ketoglutarate; Asp to oxaloacetate; Glu to a-
ketoglutarate.
? Asn is converted to Asp via a deamination reaction
catalyzed by asparaginase.
? Gln is converted to Glu also via a deamination
reaction catalyzed by glutaminase,
16,Gly can be converted to Ser or
CO2 and NH4+ for oxidation in
different organisms?
In animals,Gly is oxidized to form CO2 and NH4+,
with electrons collected by NAD+ and one carbon
unit transferred to H4 folate (to form N5,N10-
methylene-H4 folate) in a reaction catalyzed by
glycine synthase.
? In microorganisms,Gly is first converted to Ser by
accepting a hydroxymethyl group from N5,N10-
methylene-H4 folate in a reaction catalyzed by serine
hydroxymethyl transferase; Ser is then converted to
pyruvate for further oxidation.
Predominant
in animalsPredominant in bacteria
Two metabolic fates of glycine
17,Acetyl-CoA is formed from the
degradation of many amino acids
? Trp,Ala,Ser,Gly and Cys,are converted to
pyruvate first before being converted to acetyl-CoA.
? Portions of the carbon skeletons of Trp,Phe,Try,
Lys,Leu,and Ile are converted (via acetoacetyl-CoA
or directly)to acetyl-CoA.
? Some of the final steps in the degradation of Leu,
Lys,and Trp resemble steps in fatty acid oxidation.
? Tryptophan degradation is the most complex.
? Intermediates of tryptophan degradation are
precursors for synthesizing other biomolecules,
including nicotinate,serotonin,and indoleacetate.
Trp,Ala,Ser,Gly and Cys are converted to
pyruvate,before being converted to acetyl-CoA
Trp,Phe,Try,
Lys,Leu,and Ile
are also converted
to acetyl-CoA
Trp is an important
biosynthetic
precursor
18,O2 is used to break the aromatic
rings of Phe and Tyr
? Phe is converted to Tyr by hydroxylation,in a
reaction catalyzed by phenylalanine hydroxylase,a
mixed-function oxygenase (or monooxygenase)
using tetrahydrobiopterin (a pterin derivative) as the
reductant.
? Mixed-function oxygenase catalyzes the
incorporation of the two oxygen atoms of O2 into
two different products (usually with one being H2O).
? Participating the conversion are also two
dioxygenases,which catalyze the incorporation of
both oxygen atoms of O2 into one product.
? Phe and Tyr are eventually converted to
fumarate and acetoacetyl-CoA.
Action of a dioxygenase,p-hydroxyphenyl-
pyruvate is converted to homogentisate (尿黑酸 ）
via a peracid and an epoxide intermediate; both
atoms of O2 emerge in homogentisate;
19,A few genetic diseases are related to
defects of Phe catabolism enzymes
? Defects in enzymes of amino acid degradation lead
to accumulation of specific intermediates,many of
which cause diseases.
? Deficiency of phenylalanine hydroxylase causes
phenylketonuria (PKU,苯丙酮尿症 ),one of the
earliest human genetic defects of metabolism (first
revealed in 1934 in testing mentally retarded
patients,whose urine turned olive-green on addition
of FeCl3),
? The accumulated Phe is converted to
phenylpyruvate (via an aminotransferase) which
caused the color change or urine.
? Another disease related to Phe/Tyr catabolism and
of historical interest is alkaptonuria (尿黑酸症 ).
? Garrod discovered that the condition is transmitted
as a single recessive inheritable trait and could be
traced to the absence of a single enzyme (early
1900s),thus hinted the direct relation between genes
and enzymes
? He recognized that homogentisate (尿黑酸 ),the
compound that is excreted in large amount and
whose oxidation turns the urine black,is a normal
intermediate in the degradation of Phe and Tyr;
? Homogentisate dioxygenase was found to be
defective in such patients.
Phenylalanine that is
accumulated in PKU
patients is
converted to other
alternative products.
20,Leu,Ile,and Val are degraded via
reactions similar to fatty acid oxidation
? The same branched-chain aminotransferase and
branched-chain a-keto acid dehydrogenase complex
catalyzes the first two degradative reactions
(transamination and oxidative decarboxylation) of
all these three amino acids in extrahepatic tissues.
? The a-keto acid dehydrogenase complex has a
similar structure and catalyzes essentially the same
type of reaction (oxidative decarboxylation of a-
keto acids) as pyruvate dehydrogenase complex and
the a-ketoglutarate dehydrogenase complex.
? The subsequent degradation of the a-keto acids
share similar strategies of fatty acid oxidation.
? Leu is finally converted to acetyl-CoA and
acetoacetate; Val to propionyl-CoA; Ile to acetyl-
CoA and propionyl-CoA.
? The propionyl-CoA produced is converted to
succinyl-CoA via carboxylation and intramolecular
rearrangement involving free radicals using
coenzyme B12 (as previously described).
? Defect of branched-chain a-keto acid
dehydrogenase complex leads to maple syrup urine
disease,all three a-keto acids and amino acids are
accumulated and the urine of the patients smell like
maple syrup (槭糖浆 ).
Summary
? Amino acid in excess can neither be stored,nor
excreted,but oxidized or converted.
? The amino groups and carbon skeletons of amino
acids take separate but interconnected pathways.
? Liver is the major site of amino acid degradation in
vertebrates.
? PLP facilitates the transaminatin and other
transformations of amino acids.
? Glutamate collects and delivers free ammonia to the
liver.
? Gln and Glu releases NH4+ in liver mitochondria.
? NH4+ in hepatocytes is converted into urea through
the urea cycle in most terrestrial vertebrates for
excretion.
? The conversion of ammonia to urea takes five (six)
enzymatic steps.
? The rate of urea synthesis is controlled at two levels.
? The carbon skeletons of the amino acids are first
converted (funneled) into seven major metabolic
intermediates.
? Some amino acids are converted to intermediates of
citric acid cycle by simple removal of the amino
groups.
? Acetyl-CoA is formed from the degradation of many
amino acids.
? O2 is used to break the aromatic rings of Phe and
Tyr.
? A few genetic diseases are related to defects of Phe
catabolism enzymes.
? Leu,Ile,and Val are degraded via reactions similar
to fatty acid oxidation
References
? Fitzpatrick,P,F,(1999),Tetrahydropterin-
dependent amino acid hydroxylases” Annu,Rev,
Biochem,68:355-382.
? Kirsch,J,F…,And Christen,P,(1984),Mechanism
of action of aspartate aminotransferase proposed on
the basis of its spatial structure” J,Mol,Biol,
174:497-525.
? Trchinsky,Y,M,(1989),Transamination,Its
discovery,biological and chemical aspects” Trends
Biochem,Sci,12:115-117.
? Halpern,J,(1985),Mechanisms of coenzyme B12-
dependent rearrangements” Science 227:869-875.
? McPhalen,C,A….,and Chothia,C,(1992),Domain
closure in mitochondrial aspartate aminotransferases”
J,Mol,Biol,227:197-213.
