AMINO ACID

METABOLISM I
2012

József Tőzsér
E-mail: tozser@med.unideb.hu
Amino acids, as building blocks of proteins and precursors of many
other compounds are fundamental for living organisms.
The essential or nonessential feature of an amino acid is important
in biochemical and physiological aspects.
The degradation of amino acids is much more complex than those of
carbohydrates and lipids, but there are many common steps.
The degradation of amino acids occur through pathways, determined
by the end product.
The products of the pathways also determine the glycogenic and
ketogenic nature of a given amino acid.

Common features of protein building amino acids:

R1 L-configuration (except the achiral glycine)

α-amino acids (except the imino acid proline)

+
HC NH3
-
COO
METABOLIC RELATIONSHIPS OF AMINO ACIDS
There is a dynamic intracellular pool of the amino acids

70-100 g/day Gln NH4+

Urea
Exogenous source
dietary proteins Fats, sterol derivatives
300-500 g/day NH4+
Endogenous source Acetyl CoA

amino acids CO2, H2O

body proteins
energy

pyruvate or
citric acid cycle
Coenzymes (e.g. NMN)
intermediates
Neurotransmitters
Phospholipids
Porphyrins
Purines
glucose glycogen
Pyrimidines
Other nitrogenous compouns (e.g. carnithine)
ESSENTIAL AND NONESSENTIAL AMINO ACIDS
Nonessential amino acids are synthesized from metabolic precursors
in the body. Essential amino acids are those that must be supplied
in the diet
Classification is on the basis of rat and human experiments.
The essentiality of certain amino acids is conditional,
it depends on the circumstances:
• Arginine is synthesized by mammalian tissues,
but the rate is insufficient to meet the need during growth.
• Cysteine can be synthesized from Methionine, but if [Met] is
low in the diet, cysteine must be supplied.
• Tyrosine can be synthesized from Phenylalanine, but if [Phe] is
low in the diet, tyrosine must be supplied.
• Histidine is essential for infants
The following amino acids are essential (or conditionally essential):
aromatic amino acids: Phe, (Tyr), Trp
branched-chain amino acids: Leu, Val, Ile, Thr
sulfur-containing amino acids: Met, (Cys)
basic amino acids: Lys, (His), (Arg),
Proteins of the foodstuffs are not equally useful!
 PROTEINS AS ENERGY SOURCES: COMPARISON WITH
THE CARBOHYDRATE AND LIPID METABOLISM
glycogen triacylglycerols proteins

glucose glycerol + FA amino acids

pyruvate Ac-CoA

lactate oxaloacetate

citrate
glucogenic aa.
ketogenic aa. fumarate
α-KG

propionyl CoA Suc-CoA

pathway

degradation of ketogenic amino acids requires mitochondria and oxigen!

 CARBOHYDRATE, LIPID AND AMINO ACID
METABOLISMS
Carbohydrates Lipids Amino acids

central intermediate: glucose fatty acids and -

glycerol

central pathways: glycolysis lipolysis

glycogenolysis β-oxidation -
glycogen synthesis fatty acid and
gluconeogenesis TAG synthesis

storage: gycogen fat (proteins)

(limited) (unlimited) no „storage”
proteins)

essentiality: - polyunsaturated about half of

fatty acids them
(a few)
EXOGENOUS AMINO ACID SOURCES:
DIGESTION OF FOOD PROTEINS
Significance of proteins in nutrition: they are essential components

Nitrogen balance: Nin – Nout: ~ 0, positive or negative

positive: growth, pregnancy, lactation, recovery from metabolic stress

negative: metabolic stress, long term starvation, long term lack of an essential
amino acid from the foodstuff
~ 0 in healthy adults

food proteins

Nin

body proteins amino acid pool excretion

Urea
NH4+ Nout
undigested proteins
DIGESTION OF FOOD PROTEINS
Proteins are digested and amino acids, di- and
tripeptides are absorbed (exception: IgG uptake after birth)

Average daily intake: 70-100 g.

Absorbed: 170-200 g.
The extra amount comes from the sloughed-off
cells and from digestive enzymes

Absorption: predominantly Na+ cotransport in the form of

amino acids, di- and tripeptides

The absorbed di- és tripeptides are hydrolyzed intracellularly

in the enterocytes
 low pH, protein denaturation proteins
Stomach autoactivation
pepsinogen pepsin
enteropeptidase
large protein fragments
Duodenum trypsinogen trypsin

chymotrypsinogen chymotrypsin
proelastase elastase
Intestine
procarboxypeptidase carboxypeptidase

oligopeptides (60%) amino acids (40%)

endopeptidases
aminopeptidases
dipeptidases
dipeptides amino acids
Enterocytes tripeptides

Capillaries amino acids

ENDOGENOUS AMINO ACID SOURCES:
INTRACELLULAR PROTEIN BREAKDOWN
Lysosomal protein breakdown:

Characteristics: acidic pH. (~5). Lysosomes are full of various hydrolyses

(proteases, nucleases, polysaccharidases) more than 40 hydrolyses, protein
degradation to amino acids

Significance: degradation of extra- and intracellular debris, phagocytosed

microorganisms, nutrition

Ubiquitin-dependent protein breakdown

Characteristics: neutral pH, ATP-dependent degradation in proteasomes

Significance: digestion of proteins programmed for rapid destruction

digestion of misfolded, denatured, abnormal proteins
AMINO ACID TRANSPORT
Group specific transport:
Carrier mediated, energy-dependent transports.
More than 20 epithelial amino acid transporters are known!

Several brush border transport systems.

Na+-dependent and -independent systems.
Na+-cotransport: energy is utilized to pump out the ion.
Examples of amino acid transporters:

Transporter Specificity Mechanism Related disease

SLC1A1 Asp, Glu Na+ cotransport
SLC3A1/SLC7A9 Lys, Arg, Cystine Na+ independent Cystinuria
LAT1 Phe, Tyr, Met, Val Na+ independent Hartnup disease.
(SLC3A2/SLC7A5) Leu, Ile, Trp
SLC1A5 Ala, Ser,Cys Na+ cotransport

Hartnup disease: neutral aminoaciduria. Lack of luminal absorption,

kidney resorption. Symptoms are similar to essential amino acid deficiency.

As LAT1 transports several essential amino acid into the cells, if [Phe]
increases (e. g. in PKU), it blocks the transport of the others.
MODEL FOR HETEROMERIC AMINO ACID
TRANSPORTERS Heavy chain (e.g. h4F2hc)
Light chain (e.g. CD98)

Heavy chain subunit (1 TM domain), responsible for the location is

disulfide-bridged to the light chain subunit (12 TM domains) that is
responsible for the specificity.
 γ-Glutamyl cycle: active at the sites of intensive transport, e.g. in renal and
intestinal brush borders. It transports mainly Cys and Gln.

It is an example of de novo peptide synthesis

+H
3N CH COO-
R

γ-Glutamyl transpeptidase

COO-
+H
Glutathione 3N CH
γ-Glutamyl-amino acid
ADP+Pi Glutathione CH2
synthetase
ATP CH2
glycine Cys-Gly
γ-Glutamylcysteine CO NH CH COO-
γ-Glutamylcysteine
ADP+Pi R
syntethase
ATP
Cysteine γ-Glutamyl-
cyclotransferase
Glutamate +H
3N CH COO-
Oxoprolinase
ADP+Pi R
O N COO-
H
ATP 5-Oxoproline
(Pyroglutamate)
 Enzyme deficiencies of the γ-glutamyl cycle:
1. Glutathione synthetase deficiency: oxoprolinuria, aminoaciduria,
hemolytic anemia, mental retardation. Acidosis due to oxoprolinemia
+H
3N CH COO-
Alternative γ-glutamyl cycle
R

γ-Glutamyl transpeptidase

COO-
Glutathione ↓ +H
3N CH
ADP+Pi CH2
γ-Glutamylcysteine ↑↑
ATP X CH2

γ-Glutamylcysteine ↑↑ CO NH CH COO-
ADP+Pi R
γ-Glutamyl-
ATP Cysteine cyclotransferase
Glutamate
O N COO-
ADP+Pi H
5-Oxoproline
(Pyroglutamate)↑↑
ATP
 Enzyme deficiencies of the γ-glutamyl cycle:
2. γ-glutamyl cysteine synthetase deficiency: aminoaciduria, hemolytic
anemia without acidosis (oxoprolinemia) and mental retardation.
+H
3N CH COO-
R

γ-Glutamyl transpeptidase

COO-

Glutathione ↓ +H
3N CH
ADP+Pi γ-Glutamyl-amino acid ↓
CH2

ATP CH2
glycine Cys-Gly
γ-Glutamylcysteine↓ CO NH CH COO-
γ-Glutamylcysteine
ADP+Pi R
syntethase
ATP Cysteine
Glutamate +H
3N CH COO- ↓
ADP+Pi O N R
COO-
H
ATP 5-Oxoproline
(Pyroglutamate)
STRUCTURE AND FUNCTIONS OF
GLUTATHIONE (GSH)
It is found in mM concentration inside the cells
Stabile, resistant towards proteases

γ-Glutamyl-Cysteinyl-Glycine

1. Intracellular reductant. [GSH]/[GSSG] >> 1

Glutatione peroxidase/reductase system: in number of tissues this is the major
pathway of H2O2 metabolism in many cells.

H2O2 2GSH NADP+

glutatione peroxidase glutathione reductase
2H2O GSSG NADPH + H+

Protection of membrane lipids agains oxidation.

Glutathione transhydrogenases: Thiol-disulfide exchange using GSH in
protein synthesis, protein degradation , enzyme activation and inactivation.
2. Precursor of S-substituted GSH derivatives.
Glutathione S-transferases
exogenous compounds: mercapturate formation (N-acetylated,
S-substituted Cys derivatives)
endogenous compounds:
leukotriene synthesis

3. Coenzyme function: several enzymes are using GSH.

e.g. glyoxalase
cis-trans isomerization by maleylacetoacetate isomerase

4. Amino acid transport

Diagnostic importance:
γ-GT increased in liver diseases especially in obstructive jaundice.
γ -GT levels are used as a marker of alcohol induced liver disease and in liver
cirrhosis.
METABOLIC RELATIONSHIPS OF AMINO ACIDS
There is a dynamic intracellular pool of the amino acids

70-100 g/day Gln NH4+

Urea
Exogenous source
dietary proteins Fats, sterol derivatives
300-500 g/day NH4+
Endogenous source Acetyl CoA

amino acids CO2, H2O

body proteins
energy

pyruvate or
citric acid cycle
Coenzymes (e.g. NMN)
intermediates
Neurotransmitters
Phospholipids
Porphyrins
Purines
glucose glycogen
Pyrimidines
Other nitrogenous compouns (e.g. carnithine)
COMMON REACTIONS IN THE AMINO ACID
METABOLISM

1. Elimination of nitrogen:
transamination
deamination: oxidative
nonoxidative
2. Decarboxylation: oxidative
nonoxidative

3. Fate of the side chain (carbon backbone):

carboxylation
C1 transfer, transmethylation
monooxigenation
dioxigenation
( β-oxidation)
FATE OF THE NITROGEN
ELIMINATION OF THE NITROGEN FROM
THE AMINO ACIDS:
Transamination: Transaminases exist for all amino acids except Lys and Thr.
Transaminases are using pyridoxal phosphate (PLP)
as cofactor.

amino acid1 E-PLP amino acid2

ketoacid1 E-PMP ketoacid2

Most frequently used amino partners: Glu/α-KG (80%); Ala/pyruvate; Asp/oxaloacetate
Role of transamination:
usual way of strarting of amino acid degradation
funneling nitrogen into glutamate
quantitatively it is the most important reaction type
it does not eliminate the nitrogen from the amino acid pool

Classification of transaminases: enzyme family. Several enzymes (>60).

depending on localization: cytoplasmic or mitochondrial
depending on the reactants, position of the amino group (α, β, etc.)

Diagnostic importance:

ENZYME PRESENT IN

Aspartate Amino transferase (AST) Heart and Liver

Serum glutamate-oxaloacetate
transaminase (SGOT)
Alanine Amino transferase (ALT) Heart and Liver
Serum glutamate-pyruvate
transaminase (SGPT)
AMINOTRANSFERASE PROTEIN FAMILY

A superposition of the structures of four Ribbon diagram of the mitochondrial

aminotransferases taking part in amino acid branched-chain aminotransferase
biosynthesis reveals a common fold,
indicating that these proteins descended
from a common ancestor.
DEAMINATION:
In a deamination the nitrogen is removed from the amino acid pool.
Deamination could occur in an oxidative way (using reducing cofactors) or
in a nonoxidative way (with intramolecular oxidation).

In oxidative deamination a redox coenzyme (NAD(P), FAD or FMN) is used in

the reaction.

General reaction scheme of oxidative deamination:

A AH2 H2O NH4+

R R
R
C NH2+ C O
H C NH3+
COO- COO-
COO-

amino acid imino acid ketoacid

A: NAD(P)
FAD or
FMN
DEHYDROGENASES: GLUTAMATE DEHYDROGENASE (GDH)
Two isoforms (both mitochondrial):
Housekeeping form: GLUD1: liver, brain, pancreas and kidney
GLUD2: retina, testis, brain
GLUD2: in X chromosome, intronless.
Glutamate + NAD(P)+ + H2O ↔ α-Ketoglutarate + NAD(P)H + NH4+

Its role in the amino acid metabolism: transdeamination

NH4+
proteins amino acids α-KG NAD(P)H + H+

transaminases glutamate dehydrogenase

ketoacids Glu NAD(P)+

protein nitrogen → glutamate nitrogen → ammonia

Regulation: GLUD1 GLUD2

GTP allosteric inhibitor -
ADP allosteric activator allosteric activator

Leucine allosteric activator allosteric activator

HIPERINSULINISM/HYPERAMMONEMIA
(HI/HA) SYNDROME:
Mutations in exons 11 and 12 of the glutamate dehydrogenase gene.
Mutant enymes show reduced sensitivity towards GTP inhibition but retain
their ability to be activated by ADP → increased α-KG and NH3 production.
Increased α-KG stimulates
TCA cycle and oxidative
phosphorylation leading to
increased insuline production
and hypogycemia.

Mutations: S448P, H454Y, R463A.

STRUCTURE OF LIVER GDH
(GLUD1):

The enzyme is homohexameric.

Bound ligands are shown in space-

filling representations.

Glu – orange

substrate NADH – pink

ADP effector site bound NADH - brown

GTP - gray

GDH in complex with

Glu, NADH, and GTP.
 with all ligands One subunit of GDH the apo-form
antenna
domain

ADP site (NADH)

GTP

pivot helix

substrate
binding domain
Glu
substrate NADH

coenzyme binding domain

When Glu and NAD(P) binds, the coenzyme binding domain rotates around the pivot
helix to close the cleft on the ligands. GTP binding favors the closed form of the enzyme
that results tight binding of the substrate/products and hence inhibition.
HI/HA mutations are located at the N-terminal end of the pivot helix.
Oxidases: peroxisomes
D-amino acid oxidase: A = FAD. High activity on glycine and
D-amino acid substrates.
L-amino acid oxidase: A = FMN. Low activity. Role in Lys degradation.

Nonoxidative deamination
β-elimination: dehydration (Ser, Thr), desulfhydration (Cys) with PLP cofactor!!

CH 2-OH CH 2 CH 2 H2 O NH4+ CH 2
H2 O

H NH3+ H NH3 H NH2+ H O

COO- COO- COO- COO-

serine imino acid pyruvate
ammonia lyase: O O
H NH4+ H H
H C H C
N N C
H C O
_
O
_
H
N NH+ N
3
histidine urokanate
ENZYME DEFICIENCY:
Histidinemia: histidine ammonia lyase deficiency. It was discovered as
false positive for phenylketonuria screening.

It has a high frequency (1:15 000).

In many cases (but not always) concurrent with speach problems,

mental retardation.

It can be verified from skin biopsy samples.

Structure of PLP, mechanism of PLP-containing enzymes

PLP is a derivative of pyridoxine (Vitamin B6). It contains a substituted

pyridine ring.
CH2OH H O
O C
HOH2C OH _
O P OH2C OH
+ CH3 _
N O + CH3
H N
H
pyridoxine pyridoxal phosphate (PLP)

PLP is a versatile coenzyme of various enzymes in amino acid metabolism

OH H
d a b _ a. transamination
CH2 c C COO
b. decarboxylation
N
c. serine hydroxymethyl-
O H C transferase
_
O P OH2C OH d. serine dehydratase
_
(+ phosphate group in
O + CH3 glycogen phosphorylase)
N
H
MECHANISM OF TRANSAMINATION:
α-ketoacid
amino acid substrate
R
R R R -
- C COO
- C COO
-
O C COO O
H C COO H+
H+ H2O NH2
N H+ H H N
N
HC HC H H C H
HC
R' OH R' OH
OH H R' OH
+
R' + + CH3 H O +
. CH3 H N 2 N
CH3
+ CH3 N
N H H H

aldimine quinonoid ketimine pyridoxamine

intermediate phosphate
(PMP)
Steps:
- PLP is covalently attached to the enzyme via a Schiff base (imine) linkage (not shown).
- In a transimination reaction the nucleophilic amino group of the amino acid substrate
attacks the enzyme-PLP Schiff base
- Tautomerization through a resonance stabilized carbanione intermediate
- α-keto acid-PMP Schiff base is hydrolyzed to produce PMP and α-keto acid
- Regeneration of PLP occurs in the reverse way using another α-ketoacid as substrate.
How does PLP function in enzymes catalyzing different type of reactions?

Stereoelectronic control: The protein part determines the specificity of an enzyme

using PLP as cofactor.

Binding of PLP to aspartate aminotransferase:

 OH H
d a b _ a. transamination
CH2 c C COO
b. decarboxylation
N
c. serine hydroxymethyl-
O H C transferase
_
O P OH2C OH d. serine dehydratase
_
O + CH3
N
H

The bond being broken must be perpendicular to

the π-orbitals of the electron sink.
BOND ORIENTATION IN A PLP–AMINO ACID SCHIFF
BASE: THE π-ORBITAL FRAMEWORK OF A PLP–
AMINO ACID SCHIFF BASE.
FORMATION AND ELMININATION OF AMMONIA
normal level in blood: 30-60 µM
higher than 100 µM: hyperammonemic coma:
genetic or aquired
Why is ammonia toxic?
Ammonia penetrates the blood-brain barrier
Through the action of glutamate dehydrogenase it causes
α-ketoglutarate depletion
Increases Glu level that is exitotoxic
Ammonia in the astrocyte mitochondria causes ROS
production and mitochondrial damages

Sources of ammonia in the body:

Glutamate dehydrogenase: reversible enzyme. In periportal
hepatocytes it releases ammonia for the synthesis of
carbamoyl-phosphate to enter to the urea cycle

Specific deaminations: β-elimination (serine dehydratase,

desulfhydrase). Histidine ammonia lyase.
AMEN GOD
Glutaminase, asparaginase reactions H2 O NH4 +

glutamine glutamate
Intestinal bacteria also produce ammonia

Purine and pyrimidine degradation: adenylate deaminase, adenosine deaminase

cytidine and guanine deaminases
pyrimidine ring nitrogen is also a source
H2O NH4+

adenosine inosine
Purine nucleotide cycle: H2O NH4+
AMP
deaminase Aspartate
fumarate AMP IMP GTP
Adenylosuccinate
Adenylosuccinate synthetase
lyase

Adenylosuccinate GDP + Pi
Amine oxidase reactions:
O2 H2O

R-CH2-NH2 R-CHO

H2O NH4+
AMINO ACID METABOLISM OF TISSUES:
Intestine: utilizes Glu, Gln, Asn, Asp
releases NH4+, Ala, citrulline

Muscle: releases Ala (glucose-alanine cycle), Gln

(binds ammonia released from the purine nucleotide cycle)
transamination of the branched-chain amino acids

Brain: Glu, Asp, Gly: neurotransmitters

amine oxidase reactions
NH4+ elimination in the form of Gln

Liver: center of the amino acid metabolism. High Km enzymes

(amino acid tRNA synthetases: low Km)
periportal cells: glutaminase, glutamate dehydrogenase, urea cycle
perivenous cells: glutamine synthase

Kidney: glutaminase activity, NH4+ and urea secretion

Rapidly dividing cells: Gln utilization, α-KG production (anaplerosis)

Gln is also an important precursor of the nucleotide synthesis
NITROGEN TRANSPORT BETWEEN ORGANS:

brain

muscle

Glutamine
Ala

intestine
Ala Gln NH4+
liver kidney
NH4+ Urea Urea
SYNTHESIS OF CARBAMOYL-PHOSPHATE.
THE UREA CYCLE:
Nitrogen excretion of animals:
ammonotelic: ammonia excretion.
Water can immediately dilute it.
- fishes
uricotelic: uric acid excretion.
Large amount of water is not required.
- birds, ampophibians
ureotelic: urea excretion.
It appears in the evolution of amphibians
as N-elimination route.
In sharks: urea is synthesized to provide an
osmotic balance

Some aminals can shift between uric acid and

urea excretion
CARBAMOYL-PHOSPHATE SYNTHETASE TYPE I.
(CPS I.)
role: it is technically not a member of the urea cycle
it catalyses the condensation and activation of HCO3- and NH4+

localization: mitochondria
catalyzed reactions:
_
HCO3 activation Pi - NH4+ exchange carbamate activation
ATP ADP NH4+ Pi ATP ADP
O O O O O O
- _
O C OH -
O P O C OH H2N C OH H2N C O P O
- _
O O
bicarbonate carbonic acid-phosphoric acid carbamate carbamoyl-phosphate
anhydride (carbonyl-phosphate)
2ATP 2ADP +Pi
_
HCO3 + NH4+ CP
regulation: N-acetyl glutamate
allosteric activator GDH
gene level regulation: 10x, 20x Glu N-Ac-Glu x N-Ac-Orn

The cycle never stops! +

Arg
COMPARISON OF CPS I. AND CPS II.

CPS I. CPS II.

pathway: urea cycle pirimidine synthesis
localization: mitochondria cytosol
N-donor: NH4+ glutamine
activator: N-acetyl-glutamate phosphoribosyl-pyrophosphate
inhibitor: - UTP
structure: one polypeptide chain part of a multifunctional protein
STRUCTURE OF CPS II (AND CPS I)
CPS-II: This enzyme consists of two chains.
The smaller chain (yellow) contains a site for Gln
hydrolysis to generate NH4+. This domain is
largely preserved in CPS-I, although it is
catalytically inactive. This site binds N-
acetylglutamate, the allosteric activator of the
enzyme:

A catalytic site in one isozyme has been adapted

to act as an allosteric site in another isozyme
having a different physiological role.

The larger chain includes two ATP-grasp domains

(blue and red).
In one ATP-grasp domain (blue), bicarbonate is
phosphorylated to carboxyphosphate, which then
reacts with ammonia to generate carbamate.

In the other ATP-grasp domain, the carbamate is

phosphorylated to produce carbamoyl phosphate.
 + -
NH2 COO
THE UREA
-
aspartate COO
+H
NH C NH CH
3N CH
CYCLE: O CH2 CH2
CH2
NH C NH2 -
AMP +2Pi CH COO
-
COO 2

CH2 argininosuccinate
cytosol CH2
CH2
HC NH3+
citrulline
citrulline transporter CH2 ATP
COO
-

NH3+
argininosuccinate
HC argininosuccinate
-
synthetase (ASS) lyase (ASL)
COO
ornithine trans- +
carbamoylase (OTC) NH2
NH3+
O O NH C NH2
_ CH2
H2N C O P O CH2 -
COO
_ CH2
O CH2 HC
carbamoyl-phosphate CH2
arginase
CH2 CH
ornithine transporter HC NH3+ -
HC NH3+ COO
-
COO - fumarate
O COO
ornithine arginine
H2N C NH2
mitochondrion urea
REGULATION OF THE UREA CYCLE:

CPS-I:
short term regulation: activation by N-acetyl glutamate
long-term regulation: gene level regulation: high protein diet induces the
CPS-I by 10x, 20x

Urea cycle enzymes:

long-term regulation: high protein diet induces the expression of the
enzymes by 10x, 20x
THE KREBS BICYCLE:

cytosol
fumarase fumarate arginine arginase
ASL
malate urea
NAD+
MDH arginino- ornitine
succinate
NADH+ H+ carbamoyl-
oxaloacetate phosphate
GOT ASS
OTC
aspartate citrulline
CPS-I

NH4+
Glu α-KG
2 or 3 ATP
GDH
Glu
Both nitrogen could come from glutamate
mitochondrion
ENZYME DEFICIENCIES OF CP SYNTHESIS AND
THE UREA CYCLE:
Each step could be affected. Complete enzyme deficiency is not compatible
with life. Partial deficiencies: hyperammonemia, the amount of the substrate
of the blocked step is elevated.

N-acetyl-Glu synthetase deficiency: hyperammonemia,

hyperaminoacidemia, mental retardation. Therapy: low protein diet, carbamoyl-
glutamate, α-ketoacid, benzoate, phenylacetate, administration.

CPS I. deficiency: hyperammonemia, mental retardation.

Therapy: low protein diet, α-ketoacid, benzoate, phenylacetate, Arg
administration.

OTC deficiency: X-linked. Orotic aciduria. The most common deficiency of the
urea cycle.

Argininosuccinate synthetase deficiency: citrullinemia, cirtullinuria.

Argininosuccinate lyase deficiency: argininosuccinate accumulation.

Arginase deficiency: very rare, mental retardation for unknown reason.

Nongenetic enzyme deficiencies: cirrhosis, aquired hepatic coma
DETOXIFICATION REACTIONS AS ALTERNATIVE
TO THE UREA CYCLE
INTERCELLULAR GLUTAMINE CYCLE

Role of the urea synthesis in the acid-base balance:

shared function of the liver and kidney. NH4+ excretion means proton elimination.

Localization of the enzymes

of the intercellular glutamine cycle:

periportal hepatocytes:
glutaminase, GDH, CPS I.,
urea cycle enzymes

perivenous hepatocytes:
glutamine synthetase

kidney: glutaminase, GDH

A single liver lobule:

periportal cells, perivenous cells
 periportal hepatocytes perivenous hepatocytes
The cycle:

Glu
Glu

Gln urea
NH4+ Gln
NH4+

urea
NH4+ (~ 0.6 mM) urea
Gln
Gln (~ 0.3 mM) Gln

acidosis: net Gln synthesis alkalosis: net Gln uptake

DECARBOXYLATION AND CARBOXYLATION
STEPS
Oxidative decarboxylation:
α-keto acid dehydrogenase enzyme complexes:
enzyme family, mitochondrial enzymes
piruvate dehydrogenase, α-KG dehydrogenase, BCKDC
1. dehydrogenase
2. dihydrolipoyl transacylase
3. dihydrolipoyl reductase (same in the enzymes)
+ phosphorylase, phosphatase

α- KG Glu NAD NADH + H+

R R R

H C NH3+ C O C O

COO- COO- CoA-SH S-CoA

FAD
amino acid α- ketoacid TPP acyl-CoA
lipoamide
DEFICIENCY OF THE BRANCHED-CHAIN α-KETOACID
DEHYDROGENASE (BCKDC) ENZYME COMPLEX:
Maple Syrup Urine Disease (MSUD). Autosomal recessive.

Severe acidosis, mental retardation.

Any of the subunits can be affected by mutation.

Frequency: 1: 185 000 (1:176 in Old Order Mennonite population)

Thiamine administration may be helpful in some cases.

MENNO SIMONS (1496 – 1561)

THE MENNONITE MUTATION OF BCKDC

MSUD frequency is 1:176 in Old Order Mennonite population.

It is attributed to a founder effect

Tyr393 of the α subunit of E1 α2β2 heterotetramer is mutated to Asn.

Tetramerization of the E1 α2β2 is affected by the mutation.

X-RAY STRUCTURE OF THE E1 COMPONENT OF THE
HUMAN BCKDC MULTIENZYME COMPLEX:
THE α2β2 HETEROTETRAMER.
TPP

Tyr

The α/β’ interface showing the

TPP cofactor and Tyr393α which is
mutated to Asn in the Mennonite mutation
causing MSUD are shown in space filling
representations.
interaction between Tyr393α,
His385α and Asp328β’. The
Mennonite mutation impedes
tetramerization.
 RARE HEREDITARY AUTISM

Some children with autism have low blood levels of branched chain amino
acids.

Genome sequencing of six children with autism has revealed mutations in

BCKDC kinase, that stops branched-chain amino acids being depleted.

Mice lacking this gene developed neurological problems related to autism

that were reversed by dietary changes

Low blood level Val, Leu, Ile

Increased Phe, Tyr, Trp transport to brain by LAT1!
ROLE OF OXIDATIVE DECARBOXYLATION IN
THE AMINO ACID METABOLISM:
α- KG Glu

Leu, Val, Ile branched-chain α-ketoacid acil-CoA

BCKDC
transsulfuration +
transmethylation
Met α-ketobutyrate propionyl-CoA
BCKDC?
Thr dehydratase

Trp, Lys α-ketoadipate glutaryl-CoA

NONOXIDATIVE DECARBOXYLATION:

Cofactor: PLP or piruvyl group.

In piruvyl utilizing enzymes proteolytic cleavage of one subunit of the
homodimeric precursor yield Ser as N-terminal amino acid. It undergoes a
dehydration producing a pyruvyl N-terminal blocking group.

Role in the amino acid metabolism:

elimination of ω-carboxyl group of glutaryl-CoA (Lys, Trp degradation)
formation of neurotransmitters: precursor functions.