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PROTEIN AND AMINO ACID METABOLISM

AAs have nitrogen which is unstorable; any AA in excess is degraded
Catabolism: (1) removal of -amino by Transamination/oxidative deamination -Keto acid + ammonia urea or excreted; (2) carbon skeleton metabolism as CO2, water, glucose, FA, ketone bodies
Amino Acid Pool:Total AA from exogenous (dietary) and endogenous (tissue) sources; total 100g CHON; muscle 50-80%; kidney 4%, liver 10%, blood 4%
Protein Turnover:
Rate of CHON AA and AA CHON; constant if healthy person; measured as g/CHON synthesized and degraded per time
Protein degradation determined by nature, function and structure; PEST sequence = short halflives; KFERQ sequence: signal for lysosomal attack
Interorgan Amino Acid Traffic
After protein-rich meal: AA to liver circulation; free AA conc 200-250% (compared to postabsorptive state), <50% branched chain AA; liver prefers aromatic,

muscle prefers branch chain, exports valine to brain and glutamine to gut and kidney
kidney ammonia, exports alanine to liver gluconeogenesis
Postabsorptive state: maintenance of plasma AA depends on net balance between use and stores;
Muscle: 50% of free AA pool; <50% of alanine and glutamine stores
Liver: primary site of nitrogen disposal as urea; takes large alanine and glutamine stores
Kidney: serine source; takes up valine

NITROGEN METABOLISM

Removal of -amino group from AA

Transamination: major process, not for all AA; reversible transfer between AA and Keto acid (KA); both degradation and biosynthesis; catalyzed by transaminases + pyridoxal PO4 + Glu-KGlu (most common AA-KA pair)

Deamination: removal of amino group as ammonia

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Oxidative

Glutamate DH: most potent deaminator; zinc containing; NAD/NADP usage; in mitochondria; reversible, can add/subtract ammonia to Glu, source of ammonia in Urea Cycle

L-AA oxidase: with FMN FMNH2 reoxides with O2 = H2

D-amino oxidase / glycine oxidase: with FAD FADH2 reoxidises with O2 = H2O2
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Non-oxidative

Hydrolytic: Asp/Glu by Asparaginase / Glutaminase; removes amide group, glutaminase in kidney = ammonia excretes H+ as NH4 for acid-base control

Direct: histidine by histidinase = urocanic acid

Dehydrative: serine / threonine + pyridoxal phosphate = pyr / Kbutyric acid

AMMONIA DISPOSAL

Alanine Cycle exported to bloodstream; extracted by liver, deaminated pyruvate gluconeogenesis

Glutamine Cycle active in muscle and brain; Glu + NH3 + ATP Gln synthase = Gln blood kidneys / intestinal mucosal cells glutaminase Glu + MH4

Urea / Krebs-Henseleit Cycle in liver; acid-base regulation

Step
Substrate
Product
Enzyme
Action / Notes
Synthesis
NH3, CO2, 2 ATP
Carbamoyl phosphate (CP)
Mitochondrial carbamoyl phosphate synthetase I (CPS I)
CPS I allosterically stimulated by N-acteyl glutamine Glu + aCoA
Formation
CP + ornithine
Citrulline
Ornithine transcabamoylase
Citrulline to cytosol
Synthesis
Citrulline + aspartate
Argininosuccinate (ArgSuc)
Argininosuccinate synthetase
ATP AMP + PPi
Cleavage
ArgSuc
Fumarate + arginine
Agininosuccinase
Fumarate Malate for gluceneogenesis or anaplerosis (OAA); OAA aspartate for urea cycle
Cleavage
Argninine
Urea + ornithine
Argniase
Ornithine transported to mitosol

Regulation
o
Substrate Availability: ammonia = urea
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Allosteric Regulation: fine control of urea cycle: CPS I stimulated by NAG Glu + aCoA
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Enzyme Induction/Repression: enzymes in response to CHON diet or prolonged fasting
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Intermediate Concentration: intermediates may be converted to other products = conc; ornithine from Arg or Glu
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Compartmentation: mitochondrial CPS I

Hyperammonemia

Inherited Deficiencies: blood ammonia, intermediates if enzyme block; mental retardation (neurotoxic NH4 sequesters KG of TCA = ATP); convulsions, death
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Hyperammonemia I: CPS I deficiency
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Hyperammonemia II: ornithase transcarbamoylase deficiency
o
Citrullenemia: argininosuccinate synthase
o
Arginosuccinic aciduria: argnininosuccinase deficiency

Argininemia: arginase deficiency

Acquired Disorders liver inability to detoxify ammonia secondary to hepatic disorders; due to neuronal toxicity ahd hepatic coma, shift of Glu DH to Glu production depletes KG ATP

CARBON SKELETON FATES

Glucogenic Alanine, Aspartate, Methionine, Cysteine, Arginine, Histidne, Serine, Glycine, Threonine, Proline, Valine
Ketogenic Leucine, Lysine
INDIVIDUAL AMINO ACID METABOLISM
Amino Acid
Step
Substrate
Product
Transamination
Alanine
Pyruvate
Alanine

Pyruvate

Cysteine /
Cystine

Serine

OAA

Tryptophan
Aspartate
Asparagine
Glutamate
Glutamine

KG

Arginine
Proline

SucCoa

Histidine

Methionine

Valine

Enzyme
Alanine amino transferase

Reduction
Direct Oxidation then
Transamination or
Transamination
(Mercaptopyruvate) then
Transulfuration
Dehydrative Deamination
Transcarboxylation or

Cystine
cysteine
Cysteine sulfunic acid
Cysteine

Cysteine
Cysteine sulfunic acid
Pyruvate
Mercaptopyruvate

Mercaptopyruvate
Serine
Glycine

Pyruvate
Pyruvate
Serine

Metabolized (?) or

Glycine

CO2, NH3

Oxidized

Glycine

Glyoxylic acid

Serine dehydratase
Serine hydroxymethyl transferase +
N5,N10 methylene tetrahydrofolate
Mitochondrial glycine synthase,
NAD, pyridoxal PO4, THF
D-AA oxidase

Cleavage

Threonine

Glycine, Acetaldehyde

Threonine aldolase

Conversion

3C of W

Transamination

Aspartate

Alanine pyruvate via

Transamination
OAA

Hydrolysis
Transamination /
Oxidative Deamination
Hydrolysis
Cleavage
Transmination / Oxidative
Deamination
Oxidation
Spontaneous Conversion
Conversion
Direct Deamination
Conversion
Hydrolysis
Donation
Conversion
Donation
Hydrolysis
Condensation
Hydrolytic Cleavage
Dehydrative Deamination
Decarboxylation
Carboxylation
Isomerization
Remethylation

Asparagine
Glutamate

NH4, aspartate
KG

Glutamine
Arginine
Ornithine

NH3, glutamate
Ornithine, Urea
-Ketoglutarate

glutaminase

Proline
1 pyrrolene-5-carboxylic acid
Glutamic acid -semi-aldehyde
Histidine
Urocanic acid
4-imidazolone-5-proprionic acid
N-formimino group of FIGLU
Methionine, ATP
Methyl group of SAM
SAH
Homocysteine + Serine
Cystathionine
Homoserine
-Ketobutyric acid
Cysteine
Malonyl CoA
Homocysteine

1 pyrrolene-5-carboxylic acid
Glutamic acid -semi-aldehyde
KG
Urocanic acid, NH3
4-imidazolone-5-proprionic acid
N-formimino glutamic acid (FIGLU)
Glutamic acid
S-adenosyl methionine (SAM)
S-adenosyl homocysteine (SAH)
Homocysteine, adenosine
Cystathionine
Homoserine, cysteine
-Ketobutyric acid
Proprionyl CoA
Malonyl CoA
Succinyl CoA
methionine

Dehydrogenase (FAD)

Transmination, Oxidative

Valine

Succinyl CoA

Glycine

Threonine

Both Isoleucine, Phenylalanine, Tyrosine, Tryptophan

Action / Notes
Major glucogenic AA,
nitrogen transport to liver

Disorders

Cystine reductase (NADH linked)

Cystinuria (renal transport defect), Cystinosis
(cystine storage defect; renal dysfunction),

Reconverted or further
oxidized to oxalate or CO2,
H2O
Acetaldehyde oxidized to
aCoA; partly ketogenic
Partly glucogenic, rest of
molecule aCoA

Glycinuria (renal stones), 1 hyperoxaluria

(renal failure)

Aspartate transminase (AST) / Glu

OAA transaminase (GOT)
asparaginase

Argininemia (defective arginase)

Histidase lyase
Urocase

Histidinemia (pseudo+ FeCl3 test)

Formimino THF
Methionine adenosyltransferase
Homocystinuria (defective cystathione
synthase)

Cystathionine synthase
Cystathionase

Hypermethioninuria (defective methionine

adenosyl transferase),
Cystathioninuria (defective cystathionase)

Mutase, VitB12
N-methyTHF / betaine +
methycobalamine
Purely glucogenic

Maple Syrup Urine Disease/ Branch Chain

Isoleucine
Leucine
Threonine

Fumarate

Tyrosine

aCoA, acetoacetate

Phenylalanine

Decaboxylation, etc.
Dehydrative deamination
Oxidative Decarboxylation
Carboxylation
Isomerization
Transmination
Oxidation, 3C Migration,
Decarboxylation
Oxidation
Cis-Trans Isomerization
Hydrolysis
Conversion
Transamination
Reduction
Decarboxylation
Oxidative Deamination
N-acetylation
Precursor
Hydroxylation
Oxidation
Reduction
Transamination
Oxidation
N-conversion
Reduction
Oxidation
Decarboxylation
Oxidation
Hydrolysis

Acetyl CoA, Proprionyl CoA

Acetoacetate and Acetyl CoA
-Ketobutyric acid
Proprionyl CoA
Methylmalonyl CoA
Succinyl CoA
p-hydroxyphenylpyruvic acid
Homogentisic acid

Homogentisic acid
Maleylacetoacetate
fumarylacetoacetate
Acetoacetate
Tyrosine

Maleylacetoacetate
fumarylacetoacetate
Fumarate, acetoacetate
aCoA, acetate
Thiolase
p-hydroxyphenylpyruvic acid
p-hydroxyphenyllactic acid
Tyramine
p-hydroxyphenylacetic acid
N-acetyl-tyrosine
aCoA
E, NE, Dopamine, Melanin
Y
Phenylalanine hydroxylase, O2, THB
Dihydrobiopterin (DHB)
THB
DHB reductase, NADH
Phenylpyruvate
Phenylacetate
N-phneylacetylglutamine
Glutamine transferase
Phenyllactate
Kynurenine-anthranillic acid Pathway
N-formylkynurenine
W DH
kynurenin
Formylase
3-hydroxykynurenine
Alanine, 3-hydroxyanthranilate

Tyrosine
Tyrosine
Tyrosine
F
THB
DHB
Phenylalanine
Phenylpyruvate
Phenylacetate
Phenylpyruvate
W
N-formylkynurenine
kynurenin
3-hydroxykynurenine
3-hydroxyanthranilate

Tryptophan

Lysine

Isoleucine
Leucine
Threonine
-Ketobutyric acid
Proprionyl CoA
Methylmalonyl CoA
Y
p-hydroxyphenylpyruvic acid

kynurenine

Hydroxylation

kynurenine

3-hydroxykynurenine

Saccharopione Pathway
Reductive Condensation
Oxidation

K, KG, NAPDH
Saccharopine

Saccharopine
-amino adipic acid
semialdehyde, glytamic acid
-keto adipic acid

-amino adipic acid

semialdehyde

Oxidative Deamination
Cyclization to Ring

K
-keto amino caproic acid

Reduction

1 piperideine-2 carboxylic
acid
Pipecolic acid

Oxidation, Hydrolytic
Opening

Tyrosine transaminase, KG
p-hydroxyphenylpyruvate
hydroxylase, Fe2+
Homogentisic acid oxidase

2-amino benzsoylpyruvic acid

Kynurenic acid

Pipecolic Acid Pathway

-keto amino caproic acid
1 piperideine-2 carboxylic acid

Enzyme protected by VitC

Tyrosinemia tyrosinuria and phenol

aciduria; Type I: deficient fumaryl acetoacetate
hydrolase, diarrhea, vomiting, cabbage odor;
Type II: Richner-Hanhart syndrome, defective
hepatic tyrosine transaminase; tyrosine, eye
& skin lesions
Alkaptonuria deficient homogentisic acid
oxidase, darkening urine, leads ochonosis
(arthritis)
Albinism - defective tyrosinase, skin
depigmentation, melanin
Phenylketonuria Type I: deficient Phe
hydroxylase; Type II&III: deficient
dihydrobiopterin reductase; mental
retardation, depigmentation, urinary
excretion of Phe derivatives, positive FeCL3
test

Forms pyruvate, may

become nicotinic acid via
quinolinic acid
Forms acetoacetylCoA, 2AMAS may become
picolinic acid

Antagonizes excretory AA
in brain
Forms xanthurenic acid, In
extrahepatic tissue

Hartnups Disease renal and intestines

cant absorb neutral AA; deficient W-2,3dioxygenase, blood/urine W; niacin
deficiency; pellagra-like (diarrhea, dementia,
dermatitis, death)
Blue Diaper Syndrome W malabsorption,
intestinal bacteria convert unabsorbed W to
indican

Hyperlysinemia impaired K conversion to

saccharopine
DH + NAD

Reactions lead to
transamination
Can proceed to
acetoacetylCoA

Spontaneous, via Schiff

base formation

Pipecolic acid
-amino adipic acid semialdehyde

Ketoaciduria - deficient K DH; burnt urine,

death; Hypervalinemia
Methylmalonic aciduria deficient Lmethylmalonyl CoA mutase, acid secretion,
responds to VitB12

2-amino muconic acid

semialdehyde, keto adipic acid

Alternate Pathway
Deamination
Ring closure

Oxidation

Mixed
Purely ketogenic

Joins saccharopine
pathway

Catabolism

Leucine

Cleavage

HMGCoA

-hydroxy -methylglutaryl CoA

(HMGCoA)
Acetoacetate, acetylCoA

Leucine

SYNTHESIS OF NON-ESSENTIAL AA

Alanine pyruvate transamination

Aspartate OAA transamination

Glutamate KG transamination

Glutamine Glu + Glu synthase (amide formation)

Asparagine Asp + Asp synthase with Glu as N donor

Glycine Ser + transferase + THF; glyoxylic acid transamination, glycine synthase, choline
SPECIALIZED PRODUCTS FROM AA

Catecholamines (E, NE, Dopamine) - Tyrosine

Melatonin, Serotonin - Tryptophan

Thyroid Hormones - Tyrosine

Histamine - Histidine

Creatine, Creatinine Glycine, Arginine

Purely ketogenic

Serine Gly , 3-phosphoglyceric acid

Proline Glu; reversal of Pro catabolism
Cysteine Met (C skel from serine + Sulfhydryl from homocysteine)
Tyrosine Phe hydroxylation
Arginine Urea Cycle (arginino-succinase reaction)

Melanin Tyrosine
Camitine Lysine
Nicotinic Acid Tryptophan
GABA Glutamate

INBORN ERRORS OF AA METABOLISM

Usually autosomal recessive (some X-linked) = inability to metabolize a specific AA

AA-metabolic enzyme = absent/defective due to mutation and impairment of gene expression

AA pathway impairment = AA in blood (amino acidemia) or urine (amino aciduria)

Polyamines Arginine (Ornithine Putrescine Spermidine,

Spermine)

Glutathione Glutamate, Glycine, Cysteine

Carnosine, Anserine - Alanine, Histidine

Nitric Oxide Arginine

Some treated via dietary restrictions, gene therapy

Manifestations of enzymatic defects: toxic metabolite, normal metabolite, pathway diversion

HORMONAL CONTROL

Effects 2 Messenger systems, covalent modification of enzymes, enzyme induction via gene expression

Insulin anabolic to AA metabolism (countered by glucagons, E, cortisol, somatostatin, growth hormone), promotes AA uptake and synthesis in skel musc (countered by cortisol and E)

Thyroid hormone cell sensitivity to anti-insulin

Glucagon, Cortisol - AA uptake in liver, gluconeogensis, ureagenesis; induce AA degradatory enzymes and urea cycle enzymes

Disease associated with hormonal changes has profound effects on AA metabolism