amino acid oxidation and the urea cycle -...
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Amino Acid Oxidation and the Urea Cycle
Amino Acid Oxidation and the Urea Cycle
Amino Acids:Amino Acids:
• Final class of biomolecules whose oxidation contributes significantly to the generation of energy
• Final class of biomolecules whose oxidation contributes significantly to the generation of energy
• Undergo oxidation in three metabolic circumstances in animals:
• Undergo oxidation in three metabolic circumstances in animals:
During normal synthesis and degradation of proteinsDuring normal synthesis and degradation of proteins
When a diet is rich in protein; amino acids are not storedWhen a diet is rich in protein; amino acids are not stored
During starvation or in diabetes mellitusDuring starvation or in diabetes mellitus
Overview of Amino Acid Catabolism:Overview of Amino Acid Catabolism:Intracellular
proteinIntracellular
proteinDietaryproteinDietaryprotein
Amino AcidsAmino Acids
NH4
+NH4
+
ExcretionExcretion
Biosynthesis of amino acidsnucleotides, biological amines
Biosynthesis of amino acidsnucleotides, biological amines
α-keto acidα-keto acid
TCA CycleTCA
Cycle
RespirationRespiration
GlucoseGlucose
Metabolic Fates of Amino GroupsMetabolic Fates of Amino Groups
Most amino acids are metabolized in the liver
Most amino acids are metabolized in the liver
• Some of the ammonia generated is used in biosynthesis; the excess is excreted
• Some of the ammonia generated is used in biosynthesis; the excess is excreted
• Excess ammonia generated in extrahepatic tissues is transported to liver for conversion to the appropriate excreted form
• Excess ammonia generated in extrahepatic tissues is transported to liver for conversion to the appropriate excreted form
Removal of the α-amino groups occurs in the cytosol by transamination reactions
catalyzed by aminotransferases (transaminases):
Removal of the α-amino groups occurs in the cytosol by transamination reactions
catalyzed by aminotransferases (transaminases):
α-keto acid2 + amino acid
1α-keto acid
2 + amino acid
1
α-keto acid1 + amino acid
2α-keto acid
1 + amino acid
2
In liver α-keto acid1 is usually α-Kg; in muscle
it is usually pyruvateIn liver α-keto acid
1 is usually α-Kg; in muscle
it is usually pyruvate
COO-COO-
C=OC=O
CH2CH2
COO-COO-
CH2CH2
α-Kgα-Kg
++COO-COO-
H3N+H3N+
RR
CC HH
amino acidamino acid
COO-COO-
C-HC-H
CH2CH2
COO-COO-
CH2CH2
NH3+NH3+ COO-COO-
RR
C=OC=O++
glutamateglutamateα-keto acidα-keto acid
COO-COO-
C=OC=O
CH2CH2
COO-COO-
CH2CH2
α-Kgα-Kg
COO-COO-
H3N+H3N+
CH3CH3
CC HH++
alaninealanine
GPTGPT
alanine
amino-
transferase
alanine
amino-
transferase
COO-COO-
C-HC-H
CH2CH2
COO-COO-
CH2CH2
NH3+NH3+
glutamateglutamate
++
COO-COO-
CH3CH3
C=OC=O
pyruvatepyruvate
COO-COO-
C=OC=O
CH2CH2
COO-COO-
CH2CH2
α-Kgα-Kg
COO-COO-
C-HC-H
CH2CH2
COO-COO-
CH2CH2
NH3+NH3+
glutamateglutamate
++
OAAOAA
++
aspartateaspartate
COO-COO-
H3N+H3N+
CH2CH2
CC HH
COO-COO-
C=OC=OCOO-COO-
CH2CH2
COO-COO-
Aspartate aminotransferase or Glutamate-OAA transaminase (GOT)Aspartate aminotransferase or Glutamate-OAA transaminase (GOT)
Serum GPT (SGPT) and GOT (SGOT) are sensitive indicators for a number of disease
conditions.
Serum GPT (SGPT) and GOT (SGOT) are sensitive indicators for a number of disease
conditions.
• During heart attacks, damaged heart cells leak aminotransferases.
• During heart attacks, damaged heart cells leak aminotransferases.
• Damaged liver cells also leak aminotransferases. SGPT and SGOT levels are monitored in people exposed to industrial chemicals.
• Damaged liver cells also leak aminotransferases. SGPT and SGOT levels are monitored in people exposed to industrial chemicals.
The effect of transamination is to collect amino groups from many amino acids and
convert them into one, glutamate
The effect of transamination is to collect amino groups from many amino acids and
convert them into one, glutamate
• Glutamate channels amino groups into biosynthetic pathways or into reactions where nitrogenous waste products are
formed
• Glutamate channels amino groups into biosynthetic pathways or into reactions where nitrogenous waste products are
formed
How are amino groups removed from glutamate and prepared for excretion?How are amino groups removed from glutamate and prepared for excretion?
Glutamate is transported into the mitochondrial matrix where it undergoes oxidative deamination catalyzed by glutamate d’hase:
Glutamate is transported into the mitochondrial matrix where it undergoes oxidative deamination catalyzed by glutamate d’hase:
GlutamateGlutamate
NAD(P)+NAD(P)+
NAD(P)HNAD(P)H
glutamate d’haseglutamate d’haseα-Kg + NH4α-Kg + NH4
Ammonia is extremely toxic to animal tissues; it is converted to glutamine for
transport from extrahepatic tissues to the liver or kidneys.
Ammonia is extremely toxic to animal tissues; it is converted to glutamine for
transport from extrahepatic tissues to the liver or kidneys.
glutamateglutamate
COO-COO-
C-HC-H
CH2CH2
COO-COO-
CH2CH2
NH3+NH3+
NH4NH4
glutamine synthetaseglutamine synthetaseCOO-COO-
C-HC-H
CH2CH2
C=OC=O
CH2CH2
NH3+NH3+
H2NH2N
glutamineglutamine
In the liver glutamine is converted back to glutamate by glutaminase:
In the liver glutamine is converted back to glutamate by glutaminase:
COO-COO-
C-HC-H
CH2CH2
C=OC=O
CH2CH2
NH3+NH3+
H2NH2N
glutamineglutamine
NH4+NH4+
glutaminaseglutaminase
H2OH2O
UREAUREA
glutamateglutamate
COO-COO-
C-HC-H
CH2CH2
COO-COO-
CH2CH2
NH3+NH3+
• Glutamine is the major transport form of ammonia; it is present in blood in much higher concentrations than other amino acids.
• Glutamine is the major transport form of ammonia; it is present in blood in much higher concentrations than other amino acids.
• Alanine also plays a role in transport of amino groups to the liver by the glucose-alanine cycle:
• Alanine also plays a role in transport of amino groups to the liver by the glucose-alanine cycle:
AA glutamate
Muscle
glutamate
alaninepyr
+ α−Kg
Liver
alanine
α−Kg
glutamatepyr +
glucose
Habitat determines the Molecular Pathway for Nitrogen Excretion
Habitat determines the Molecular Pathway for Nitrogen Excretion
• Aquatic organisms (bacteria, protozoa, fish) release ammonia to their aqueous enviroment (ammonotelic)
• Aquatic organisms (bacteria, protozoa, fish) release ammonia to their aqueous enviroment (ammonotelic)
• Birds and reptiles convert amino nitrogen into uric acid; they cannot carry enough water for the excretion of
nitrogen as urea (uricotelic)
• Birds and reptiles convert amino nitrogen into uric acid; they cannot carry enough water for the excretion of
nitrogen as urea (uricotelic)
• Terrestrial animals excrete amino nitrogen in the form of urea (ureotelic)
• Terrestrial animals excrete amino nitrogen in the form of urea (ureotelic)
• Plants recycle virtually all amino groups; there is no general pathway for nitrogen excretion
• Plants recycle virtually all amino groups; there is no general pathway for nitrogen excretion
The Urea CycleThe Urea Cycle
• Discovered by Hans Krebs and Kurt Hanseleit
• Discovered by Hans Krebs and Kurt Hanseleit
• Occurs in the liver• Occurs in the liver
• Takes place in two intracellular compartments; the cytosol and the mitochondrial matrix
• Takes place in two intracellular compartments; the cytosol and the mitochondrial matrix
NH4+NH4+ HCO3
-HCO3-++ carbamoyl phosphate
synthetase I
carbamoyl phosphate
synthetase I
2 ATP2 ATP2 ADP,
Pi2 ADP,
Pi
H2N-C-O-P-O-H2N-C-O-P-O-
OOOO
O-O-
carbamoyl phosphatecarbamoyl phosphate
ornithine transcarbamoylase
ornithine transcarbamoylase
NH3-(CH2)3-CH-COO-NH3-(CH2)3-CH-COO-
NH3+NH3+
OrnithineOrnithineH2N-C-NH-(CH2)3-CH-COO-H2N-C-NH-(CH2)3-CH-COO-
NH3+NH3+
CitrullineCitrulline
OO
PiPi
Transported to cytosolTransported to cytosol
H2N-C-NH-(CH2)3-CH-COO-H2N-C-NH-(CH2)3-CH-COO-
NH3+NH3+
CitrullineCitrulline
OO AspartateAspartate
ATPATP
PPiPPi
arginosuccinate
synthetase
arginosuccinate
synthetase
-C-NH-(CH2)3-CH-COO--C-NH-(CH2)3-CH-COO-
NH3+NH3+NH2
+NH2+
-OOC-CH2-CH-NH-OOC-CH2-CH-NHCOO-COO-
ArginosuccinateArginosuccinate
-C-NH-(CH2)3-CH-COO--C-NH-(CH2)3-CH-COO-
NH3+NH3+NH2
+NH2+
-OOC-CH2-CH-NH-OOC-CH2-CH-NHCOO-COO-
ArginosuccinateArginosuccinate
arginosuccinate lyasearginosuccinate lyase
-OOC-CH=CH-COO--OOC-CH=CH-COO-
FumarateFumarate
-C-NH-(CH2)3-CH-COO--C-NH-(CH2)3-CH-COO-
NH3+NH3+NH2
+NH2+
NH2NH2
ArginineArginine
-C-NH-(CH2)3-CH-COO--C-NH-(CH2)3-CH-COO-
NH3+NH3+NH2
+NH2+
NH2NH2
ArginineArginine
arginasearginase
NH3-(CH2)3-CH-COO-NH3-(CH2)3-CH-COO-
NH3+NH3+
OrnithineOrnithine
H2N-C-NH
2H
2N-C-NH
2
OO
UREAUREA
Back to mitochondrionBack to mitochondrion
Overall equation for Urea synthesis:Overall equation for Urea synthesis:
NH3 + HCO3- + Aspartate + 3 ATPNH3 + HCO3- + Aspartate + 3 ATP
urea + fumarate + 2ADP + 2Pi + AMP + PPiurea + fumarate + 2ADP + 2Pi + AMP + PPi
Regulation of the Urea Cycle:Regulation of the Urea Cycle:
• Carbamoyl Phosphate Synthetase I is allosterically activated by
N-acetylglutamate.
• Carbamoyl Phosphate Synthetase I is allosterically activated by
N-acetylglutamate.
• High levels of transamination during amino acid breakdown lead to elevated glutamate with concommitant increases in the concentration of N-acetylglutamate .
• High levels of transamination during amino acid breakdown lead to elevated glutamate with concommitant increases in the concentration of N-acetylglutamate .
Breakdown of Individual Amino AcidsBreakdown of Individual Amino Acids
Degradation of the carbon skeletons of the 20 common amino acids yields one of 7
intermediates: a-Kg, succinyl CoA, pyruvate, fumarate, OAA, acetoacetate, acetyl CoA
Degradation of the carbon skeletons of the 20 common amino acids yields one of 7
intermediates: a-Kg, succinyl CoA, pyruvate, fumarate, OAA, acetoacetate, acetyl CoA
α-Kg, succinyl CoA, pyruvate, fumarate, OAA can all serve as precursors for glucose synthesis; hence amino acids giving rise to
these intermediates are Glucogenic.
α-Kg, succinyl CoA, pyruvate, fumarate, OAA can all serve as precursors for glucose synthesis; hence amino acids giving rise to
these intermediates are Glucogenic.
Acetoacetate and acetyl CoA can serve as precursors for fatty acid or ketone
synthesis; hence amino acids giving rise to these compounds are termed Ketogenic
Acetoacetate and acetyl CoA can serve as precursors for fatty acid or ketone
synthesis; hence amino acids giving rise to these compounds are termed Ketogenic
Alanine, Serine, Glycine,
Cysteine, Threonine,
Tryptophan
Alanine, Serine, Glycine,
Cysteine, Threonine,
Tryptophan PyruvatePyruvate
Isoleucine, Leucine,
Threonine, Tryptophan
Isoleucine, Leucine,
Threonine, Tryptophan Acetyl CoAAcetyl CoA
Leucine, Lysine,
Phenylalanine,
Tyrosine
Leucine, Lysine,
Phenylalanine,
Tyrosine
AcetoacetateAcetoacetate
Arginine, Glutamine,
Glutamate, Proline, Histidine
Arginine, Glutamine,
Glutamate, Proline, Histidine α−Kgα−Kg
Isoleucine, Methionine,
Valine
Isoleucine, Methionine,
ValineSuccinyl CoASuccinyl CoA
Aspartate, Tyrosine,
Phenylalanine
Aspartate, Tyrosine,
PhenylalanineFumarateFumarate
Aspartate,
Asparagine
Aspartate,
AsparagineOAAOAA
ThreonineThreonineH3C-C-C-COO-H3C-C-C-COO-
HH
NH2NH2HOHO
HH
C-COO-C-COO-
NH2NH2
HHHH
GlycineGlycine
C-COO-C-COO-
NH2NH2
HOCH2HOCH2
HH
SerineSerineCH3-C-COO-CH3-C-COO-
OO
PyruvatePyruvate
TryptophanTryptophan
C-COO-C-COO-
NH2NH2
CH2CH2
HH
NHNH
C-COO-C-COO-
NH2NH2
CH3CH3
HHAlanineAlanine
C-COO-C-COO-
NH2NH2
HS-CH2HS-CH2
HH
CysteineCysteine
• Alanine is converted to pyruvate by transamination.• Alanine is converted to pyruvate by transamination.
• Asparagine is converted to Aspartate by Asparaginase:• Asparagine is converted to Aspartate by Asparaginase:
C-COO-C-COO-
NH2NH2
C-CH2C-CH2
HHOO
NH2NH2
C-COO-C-COO-
NH2NH2
C-CH2C-CH2
HHOO
-O-O
• Aspartate can be converted to OAA by transamination:• Aspartate can be converted to OAA by transamination:
Aspartate + α-KG Glutamate + OAAAspartate + α-KG Glutamate + OAA
• Aspartate degradation via the urea cycle yields fumarate.• Aspartate degradation via the urea cycle yields fumarate.
Methionine, Valine, IsoleucineMethionine, Valine, Isoleucine
Propionyl CoAPropionyl CoA
CH3-CH2-C-S-CoACH3-CH2-C-S-CoAOO
-OOC-CH2-CH2-C-S-CoA-OOC-CH2-CH2-C-S-CoAOO
Succinyl CoASuccinyl CoA
Arginine, Glutamine, Histidine, Proline Arginine, Glutamine, Histidine, Proline
GlutamateGlutamate
Glutamate D’haseGlutamate D’hase
α−Kgα−Kg
Leucine and Lysine are the only two purely ketogenic amino acids. HMG-CoA is an
intermediate in leucine degradation.
Leucine and Lysine are the only two purely ketogenic amino acids. HMG-CoA is an
intermediate in leucine degradation.
• Initial steps in valine, leucine, isoleucine degradation are identical:
• Initial steps in valine, leucine, isoleucine degradation are identical:
transamination to the corresponding a-keto acidtransamination to the corresponding a-keto aciddecarboxylation to the corresponding CoA derivativedecarboxylation to the corresponding CoA derivativedehydrogenation to form a double bonddehydrogenation to form a double bond
Defect in the decarboxylation reaction results in Maple Syrup Urine Disease, which is fatal unless treated early in life.
Defect in the decarboxylation reaction results in Maple Syrup Urine Disease, which is fatal unless treated early in life.
Phenylalanine is converted to Tyrosine by Phenylalanine Hydroxylase; then the pathway proceeds with the breakdown of tyrosine to fumarate plus acetoacetate. Homogentisate is an intermediate in this pathway.
Phenylalanine is converted to Tyrosine by Phenylalanine Hydroxylase; then the pathway proceeds with the breakdown of tyrosine to fumarate plus acetoacetate. Homogentisate is an intermediate in this pathway.
Alkaptonuria results in the urinary excretion of excess homogentisate; air oxidation causes this compound to turn dark. This disease is not fatal, individuals tend to suffer arthritis in later life.
Alkaptonuria results in the urinary excretion of excess homogentisate; air oxidation causes this compound to turn dark. This disease is not fatal, individuals tend to suffer arthritis in later life.
OHOH
OHOH-OOC-H
2C-OOC-H
2C homogentisatehomogentisate
Phenylketonuria results from absence of phenylalanine hydroxylase. Phe is converted to phenylpyruvate and excreted. Severe mental retardation occurs unless infants are immediately placed on a diet low in Phe.
Phenylketonuria results from absence of phenylalanine hydroxylase. Phe is converted to phenylpyruvate and excreted. Severe mental retardation occurs unless infants are immediately placed on a diet low in Phe.
CH2
CH2
C=OC=O
COO-COO-
phenylpyruvatephenylpyruvate
Human Genetic Disorders of Amino Acid Catabolism
Human Genetic Disorders of Amino Acid Catabolism
ConditionConditionIncidenceIncidence
(per 100,000 births)(per 100,000 births) Defective EnzymeDefective Enzyme SymptomsSymptoms
AlkaptonuriaAlkaptonuria 0.40.4 Homogentisate DioxygenaseHomogentisate Dioxygenase
Dark pigment in urine; arthritis in late lifeDark pigment in urine; arthritis in late life
Maple Syrup Urine DiseaseMaple Syrup Urine Disease
0.40.4 Branched chain α-keto acid dedydrogenaseBranched chain α-keto acid dedydrogenase
Mental retardation; convulsions; early death
Mental retardation; convulsions; early death
PhenylketonuriaPhenylketonuria 88 phenylalanine hydroxylasephenylalanine hydroxylase
Neonatal vomiting; mental retardationNeonatal vomiting; mental retardation
Methylmalonic Acidemia (MMA)Methylmalonic Acidemia (MMA)
(<0.5)(<0.5) Methylmalonyl CoA Mutase Methylmalonyl CoA Mutase
Mental retardation; convulsions; early death
Mental retardation; convulsions; early death