AMINO ACID

METABOLISM III
2012

József Tőzsér
E-mail: [email protected]
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
(-oxidation)
MONOOXYGENATION AND DIOXYGENATION
In monooxygenation one atom of molecular oxygen forms hydroxyl
group of the substrate while the other forms water with the
hydrogens of the cofactor

Role of monooxygenation (hydroxylation) in amino acid metabolism:

Phe degradation, neurotransmitter synthesis from Tyr, Trp:
Aromatic amino acid hydroxylase enzyme family

Common characteristics of the enyme family: H H

H2N N N H
C
Cofactor: tetrahidrobiotrerine (BH4), H
N C
it is not vitamine! C N C CH CH3
H
Catalytic domain: high homology, O OH OH
the substrate binding sites are different
tetrahydrobiopterine (BH4)
Regulatory domain: little homology
Phe hydroxylase is regulated by phosphorylation
(turning off the enzyme)

NH2- substrate specificity, regulation conserved domain: BH4 binding, active center -COO-
 Example: phenylalanine hydroxylase
(liver, kidney)
NADPH + H+
O
dihidrofolate
reductase
O BH2 synthesis
+ NADP+ from GTP
NH 3
phenylalanine

O2 BH4 NAD+

phenylalanine dihydrobiopterine
hydroxylase reductase

H2O
qBH2 NADH + H+
O

Spontaneous, small process

O
+
NH 3
OH
tyrosine
PTERINS ARE REDOX COFACTORS

Pterins are compounds that contain

the pterin ring. The pterin ring
resembles to the isoalloxazine
ring of flavin coenzymes.

Pterins like flavins participate in

redox reactions

Biopterin and folate contain

pterin rings, and they act in
the tetrahydro form. Folate is
a vitamin while biopterin is
synthesized in the body
ENZYME DEFICIENCIES OF PHETYR
CONVERSION:
Phenylalanine hydroxylase deficiency: classical phenylketonuria (PKU).
Most common desease of the amino acid metabolism. Autosomal recessive.
Mental retardation, light skin color.
Therapy: synthetic diet low in phenylalanine.

Dihydrobiopterine reductase deficiency: only 3 % of PKU.

Tyr and Trp hydroxylation is also deficient  central nervous functions are
more seriously affected.
Therapy involves 5-hydroxytryptophan and DOPA administration.

Maternal PKU: Even if the neonate is a heterozygote, high Phe concentration

in the mother's blood causes severe mental retardation and heart problems
of the fetus.

Enyzmes of the BH2 synthesis: In very rare cases the synthesis of the
cofactor from GTP may also be affected due to the deficiency of one of
the enzymes catalyzing the process.
STRUCTURE OF THE PHENYLALANINE
HYDROXYLASE (PAH).
PAH is a non-heme iron-containing enzyme thai is active in a
tetrameric form. It contains Fe2+ in its active center, which is not
part of iron-sulfur cluster.

Mutations in the genes encoding this enzyme

cause classical PKU.
More than 400 PKU mutations have been
Identifie.

PKU mutations include mutations of the

active site (blue)
biopterin-binding site (red),
other regions (purple)

NH2- regulatory domain catalytic domain tetramerization domain -COO-

COMMON PKU MUTATIONS IN EUROPE.

Regional distributions, possible origins and migration routes are shown.

DIOXYGENATION:
both atoms of molecular oxygen enters the substrate
Role: opening of aromatic rings
in the degradation of Phe and Tyr:

OH O
O homogentisate
O
O dioxygenase O

O Fe 2+, vitamin C O

OH O
O O

homogentisate maleylacetoacetate
ALKAPTONURIA:
• Disease with several symptoms. Ochronosis, urine turns black when
exposed to air.
• In 1898, an English doctor named Archibald Garrod showed that the
substance responsible of the urine blackening is homogentisic acid.
• In 1902, early in the post-Mendelian era, Garrod suggested, on the
basis of pedigree patterns, that alkaptonuria is inherited as a
Mendelian recessive.
• In 1908, he proposed that the disorder was caused by the lack of an
enzyme: it is among the earliest proposed cases of an "inborn error
of metabolism”
• In 1978 Stenn and his coworkers reported that an Egyptian mummy,
dating from 1500 B.C., was alkaptonuric, based on Roentgenograms
and chemical analysis of biopsy samples. This is, so far as known,
the earliest verified case of this disorder.
URINE SAMPLE OF ALKAPTONURIC PATIENT

+ NaOH

Eye of an alkaptoruric patient:

DIOXYGENATION IN TRP DEGRADATION:

O O
tryptophan
O O dioxygenase O
+
NH3
+
O NH 3
O N
N Fe 2+, HEM
formylkynurenine
tryptophan
CH
O
COO
COO 3-hidroxy-anthranilate
dioxygenase
O
O
O NH2
NH2
O O
OH
3-hidroxy-anthranilate 2-amino-3-carboxymuconic
semialdehyde
FATE OF THE SKELETONS OF AMINO ACIDS
The degradation of amino acids provides about 10-15% of the required energy
in the human body. The carbon skeletons of the 20 amino acids are funneled
into seven molecules: pyruvate, Ac-CoA, AcAc-CoA, α-KG, Suc-CoA, fumarate
and oxaloacetate.
Leu, Lys
Ala, Thr Phe, Tyr
Cys, Gly Leu, Ile Trp
pyruvate
Ser, (Trp)

Asp, Asn Acetoacetyl-CoA

Acetyl-CoA
COLOR CODING:
oxaloacetate - glucogenic amino acid
Phe
- ketogenic amino acid
Tyr fumarate
- both glucogenic and ketogenic
citrate
Ile, Met
Thr, Val Suc-CoA
-KG
Glu
Arg, Gln
His, Pro
 Leu, Lys
Ala, Thr Phe, Tyr
Cys, Gly Leu, Ile Trp
pyruvate
Ser, (Trp)

Asp, Asn Acetoacetyl-CoA

Acetyl-CoA

oxaloacetate Most of the essential amino acids

Phe are degraded on these pathways:
Tyr fumarate fatty acid-like degradation
citrate
Ile, Met
Thr, Val Suc-CoA
-KG
Glu
Arg, Gln
His, Pro

Ketogenic amino acids: they are converted to Acetyl-CoA or Acetoacetyl-CoA. They

cannot be utilized for anaplerosis and gluconeogenesis. Purely ketogenic: Leu, Lys.
Glucogenic amino acids: their degradation yield pyruvate or citric acid intermediate,
therefore they can be precursors for gluconeogenesis. (However, they can also be
converted to Ac-CoA!)
PYRUVATE PATHWAY
General characteristics: Mainly glucogenic, nonessential amino acids.

Trp: also ketogenic (AcAc-CoA pathway)

Thr: essential, also degrades through Suc-CoA pathway
Ser: synthesis from 3-phosphoglycerate,
degradation depends on the energy requirements
Ser-Gly conversion is reversible
Cys: its synthesis is Met-dependent.

(Trp) Gly (Thr)

Ala Ser 3-PG

Cys

Pyruvate
DEGRADATION OF CYSTEINE

CH 3 Degradation of Cys: major

H2S NH4+ C O source of sulfur for the body
- X = CN-
COO
H2O PLP sulfur transfer. RSH
pyruvate GSH
SO3 2-
CH 2-SH -KG Glu CH 2-SH X S-X CH 3
H C NH3+ C O C O
- - -
COO PLP COO COO
-mercapto-pyruvate pyruvate
cysteine
O2 - 2-
CH 2- SO 2 -KG Glu CH2SO2- H2O HSO3 CH 3
+
H C NH 3 C O C O
- - -
COO COO COO
cysteinesulfinate sulfinylpyruvate pyruvate
CO2 PLP
- ½ O2 -
CH 2- SO 2 CH 2- SO 3
+ +
H C NH 3 H C NH 3
H2 H2
hypotaurine taurine
FATE OF SULFITE, STRUCTURE, SYNTHESIS AND
UTILIZATION OF PAPS:

H 2O H 2 O2

HSO3- SO42- + H+ adenine

O2 ATP
O O O
ATP sulfurase PPi ATP ADP
- -
O S O P P O
AMPS phosphokinase - -
-
O O O
adenosine phosphosulfate
(AMPS) phosphoadenosine phosphosulfate
(PAPS)
SYNTHESIS OF CYS: METHIONINE-DEPENDENT:
TRANSMETHYLATION + TRANS-SULFURATION
Met SAM SAH homoCys

H2C SH OH SH
H 2C S CH2 CH3
CH2 CH2 H 2O H 2O NH 4+
PLP CH2 + PLP CH2 CH2
HC NH3
+
+ HC NH3 +
+ HC NH3 HC NH3
+
- + HC NH3
-
COO cystathionase C O
COO
-
COO cystathionine - COO
-

synthetase COO COO

-

homocysteine serine
-ketobutyrate cysteine
cystathionine

BCKDC?
transz-sulfuration: S-transfer from homocysteine to the
serine backbone
propionyl-CoA

Precursor functions of Cys: GSH

sulfuration
taurin
ENZYME DEFICIENCIES OF TRANS-
SULFURATION:
Cystathionin synthase: homocystinemia, homocistinuria, hipermethioninemia.
Homocysteine may react with and block lysyl aldehyde groups in collagen.
The lens of eye is frequently dislocated. Other ocular abnormalities often
occur. Mental retardation is frequently the first indication.
Excess homocystein may damage LDL through reactions with the amino
termini, and causing LDL aggreagtion. Aggregated LDLs are endocytosed by
macrophages. The lipid deposits form atheromas.

Cystathionase: cystathionemai, cystathionuria

Therapy: restriction of Met intake. In some cases significant improvement has

been obtained by feeding pyridoxine (vitamin B6).
FORMATION AND UTILIZATION OF C1 UNITS,
TRANSMETHYLATION
folic acid
NADPH + H+ (cystathionine)
dUMP dTMP dihydrofolate reductase (transsulfuration)
NADP+
FH2 PLP
NADPH + H+
dihydrofolate reductase
Ser NADP+ Met synthetase

FH4 methyl-B12 homoCys SAH methylated

PLP Gly X
Gly PLP
NH4+ + HCO3- betain X: norepinephrine
ethanolamine
methylene-FH4 dimethylglycine DNA, RNA
PPi + Pi guanidinoacetate
NAD+ ATP

methyl-FH4 B12 Met SAM X

NADH +H+
NH4+ EXOGENOUS methyl-FH4
methenyl-FH4 formimino-FH4 His
H2O
FH4

formyl-FH4 formate Trp

ADP +Pi ATP

Purine nucleotide (dcSAM)
synthesis (polyamines)
DEGRADATION OF SERINE

Degradation of Ser depends on whether it is used for energy or

for gluconeogenesis
serine dehydratase: uses PLP
CH2-OH H 2O CH2 NH4+ CH
CH3 H 2O 3

+ + +
HC NH3 C NH3 C NH2 C O
- - -
COO COO COO COO-
Towards gluconeogenesis:

CH2-OH -KG Glu CH2-OH NADH+H+

2-
NAD+ CH2-OH ATP ADP CH2OPO3
+
HC NH3 C O HC OH H C OH
-
transaminase -
D-glycerate glycerate kinase
- -
COO COO dehydrogenase COO COO
serine 3-hydroxypyruvate D-glycerate 3-phosphoglycerate
 SYNTHESIS OF SERINE

Synthesis of serine is from 3-phosphoglycerate instead of pyruvate:

2- 2- 2-
CH 2 OPO 3 CH2 OPO 3 CH 2 -OPO 3 CH 2 -OH
NAD + NADH+H + -KG Glu H 2O Pi
HC OH C O H C NH 3 + HC NH 3
+

-
dehydrogenase -
transaminase phosphatase
- -
COO COO COO COO
3-phosphoglycerate 3-phosphopyruvate 3-phosphoserine serine

Precursor functions of Serine: phospholipids (choline, ethanolamine)

synthesis of Cys
synthesis of Gly
Serine hydroxymethyl transferase:
C1 source
DEGRADATION OF GLYCINE

With D-amino acid oxidase:

NH4+ NADH+H+ NAD+

glycine glyoxalate oxalate

FAD FADH2
-KG
-hydroxy--ketoadipate
carboligase O
-
- COO
OOC
OH
GLYCINE SYNTHETASE: REVERSIBLE ENZYME,
MAY ALSO SYNTHESIZE GLY

FH4 methylene-FH4
PLP
lipoic acid
Gly HCO3- + NH4+
FAD
NAD+ NADH + H+

Glycine synthetase is a multienzyme

complex, resembling PDC.
It contains four proteins:
P: PLP-dependent Gly decarboxylase
H: lipoamide containing aminomethyl
carrier
T: FH4 containing aminomethyltrasferase
L: NAD+-dependent, FAD-requiring
dihydrolipoyl dehydrogenase
(E3 of PDC)
SERINE HYDROXYMETHYL TRANSFERASE:
INTERCONVERSION OF SER AND GLY

FH4 methylene-FH4
PLP
Ser Gly

Precursor functions of glycine: neurotransmitter

purine synthesis
porphyrine synthesis
creatine synthesis
bile acid
glutathione
detoxification
C1 source
SYNTHESIS AND DEGRADATION OF ALANINE

Synthesis and degradation of alanine by transamination:

alanine aminotransferases
Ala -KG
Transport of Ala: glucose-alanine cycle
pyruvate Glu
DEGRADATION OF THREONINE
Threonine is an essential amino acid that is both H3C C OH
glucogenic and ketogenic
+
Degradation of Thr: there is no transaminase. HC NH3
-
Threonine dehydrogenase pathway: the major route COO

NAD + NADH+H + CO 2 NH 4 +

Thr -amino--ketobutyrate aminoacetone methylglyoxal pyruvate

spontaneous amine oxidase oxidation
Threonine (serine) dehydratase:
NH 4+

Thr -ketobutyrate propionyl-CoA

PLP oxidative decarboxylation

Threonine aldolase (serine hydroxymethyl transferase):

the C2 unit is too big to be carried by FH4
FH4
Thr Gly + acetaldehyde Ac-CoA
PLP
α-KETOGLUTARATE PATHWAY
General characteristics:
nonessential and conditionally essential amino acids
glucogenic character
Glu: key role in NH4+ binding and elimination
His: conditionally essential, important C1 source
Orn: if any Orn is made de novo, it is made from glutamate
Pro: both synthesis and degradation occurs in this pathway

Orn Arg
Pro

glutamate semialdehyde

His Glu Gln

α- KG
DEGRADATION OF HISTIDINE

Histidine is a conditionally essential amino acid. Its degradation has a greater

importance in the C1 metabolism than in energy production. Transamination is
the first step of a minor pathway. The major degradation pathway starts with
ammonia lyase (histidase).

NH4+ formimino-FH4
FH4
H 2O H 2O

histidine urokanate formiminoglutamate glutamate

O
- H COO
-
N COO N
N -
O
COO-
N NH3 N NH2+
H
H
O OH
H

typical and special water addition steps

Measurement of FIGLU after His administration: FIGLU (formiminoglutamate) in

the urine is a measure of functional FH4 in the body.
ENZYME DEFICIENCY:
Histidinemia: histidin ammonia lyase deficiency. It was discovered as fals
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.

PRECURSOR FUNCTIONS OF HISTIDINE

Histamine in inflammation, allergic reactions

Carnosine (β-Ala-His), anserine (β-Ala-metil-His):
typically in skeletal muscle (myosine ATP-ase activation).
The are de novo synthesized from amino acids place of
methylation
(like glutathione).
β-Ala is produced as a degradation product of
O
pyrimidines
+
NH3 N -
N O
N NH
N
O
histamine
H2N

carnosine

ATP PPi His AMP SAM SAH

-Ala -Ala-adenylate -Ala-His anserine

(carnosine)
 SYNTHESIS AND DEGRADATION OF PROLINE

Proline degradation and synthesis occurs on the same pathway, however, some
reactions are catalyzed by different enzymes in different compartments.

H 2O
NAD+ NADH + H+ NAD+ NADH + H+
mitochondrion mitochondrion
H 2O

proline cyclic Schiff base -glutamyl semialdehyde Glu

90% 10%

cytosol cytosol
NADP+ NADPH + H+ NAD+ NADH + H+
O O O ATP
ADP +Pi
O
O O O
NH N NH3+
O H
H
DEGRADATION, SYNTHESIS OF ARG, ORN,
THEIR PRECURSOR FUNCTIONS.
Arginine can be converted to ornithine by arginase which can also be found
in tissues other than liver.
Ornithine transaminase: δ-aminotransferase, catalyses a reversible reaction,
however, due to the low concentration of
γ-glutamyl-semialdehyde, only low amount of Orn Arg
Pro
Orn and Arg can be synthesized. Orn can be 10%
converted to Arg by the enzymes of the Urea cycle glutamate semialdehyde

Synthesis of Arg in lower organisms: 90%

His Glu Gln
AcCoA CoA-SH

α- KG
Glu N-Acetyl-Glu N-Acetyl-Orn Orn
In mammals only the first step exists (see activation of CPS I.)
Precursor functions:
Synthesis of nitric oxide (NO), a gas phase neurotransmitter from arginine:

NOS
Arg citrulline + NO
BH4
NADPH
CREATINE
Creatine phosphate is an important reservoir of high-energy phosphate groups
in skeletal muscle.
Its phosphate can be transferred to ADP with the help of creatine kinase:

ATP ADP
creatine
kinase

creatine phosphocreatine

ATP ADP

Creatine kinase occurs as a dimer. There are two types of subunits,

M (muscle type) and B (brain type). The isoenzyme containing both type of
subunits (MB) is found only in myocardium.
SYNTHESIS OF CREATINE
NH2 NH2
+
+
C NH2 C NH2
+ +
NH3 NH NH NH3
+ CH2 CH2 CH2
CH2 +
-
-
CH2 COO CH2
COO guanidinoacetate
CH2 CH2
+ SAM +
HC NH3 HC NH3
glycine - -
COO COO
SAH
arginine +
ornithine
NH2
-
NH2 C N CH2 COO
CH3
creatine
Creatinine formation: spontaneous loss of the high enery bond: "cost of storage„
+ O
O NH2 NH
C
-
O P NH2 C N CH2 COO HN C + Pi
CH2
O CH3 N
CH3

phosphocreatine creatinine
POLYAMINES:
PUTRESCINE, SPERMIDINE, SPERMINE.
Structure: they contain multiple positive charges.
+
+ NH3
NH3
putrescine
+
+ NH3
NH3 N
H
spermidine
H +
+ N NH3
NH3 N
H
spermine
Functions: their synthesis is prerequisit for cell division.
Inhibitors of polyamine synthesis may cause terminal differentiation of some
tumors of embrionic origin.
Inhibitors of ODC also have antiparasitic effect.
Polyamines can crosslink noncovalently the double stranded DNA, and neutralize
the phosphate backbone.
SYNTHESIS OF POLYAMINES

SAMDC
SAM dcSAM
pyruvyl MTA MTA

ODC
ornithine putrescine spermidine spermine
PLP spermidine synthetase spermine synthetase
CO 2

ODC: ornithine decarboxylase. Inducible enzyme. Contains PEST sequence.

Can be inhibited by difluoro-methyl-ornithine (DFMO)

SAMDC: S-adenosyl-methionine decarboxylase. Inducible enzyme, the only

mammalian decarboxylase using pyruvyl cofactor. It produces decarboxylated SAM (dcSAM)

spermidine synthetase: constitutively expressed.

spermine synthetase: constitutively expressed.

CENTRAL ROLE OF GLU AND GLN IN THE
AMINO ACID METABOLISM

Interconversion of Gln, Glu and α-KG:

amino acid ketoacid ADP NH4+

-glutamyl-P
ATP Pi
transaminase glutamine synthetase
-KG glutamate glutamine
dehydrogenase
glutaminase

NAD(P)H + H+ NAD(P)+ H 2O
NH4+
NH4
+

Glutamine: N-transport

N-donor in some reactions: Asn synthesis, CPS II. catalyzed reaction,

some steps of purine and pyrimidine synthesis
NAD formation, aminosugar synthesis
OXALOACETATE PATHWAY:
METABOLISM OF ASPARTATE AND ASPARAGINE
These amino acids do not have such a role as Gln and Glu in
metabolism.

H2O Asn Glu + AMP + PPi

Asparaginase Asparagine synthetase

NH4+ Asp Gln + ATP

oxaloacetate

L-asparaginase is an effective chemotherapeutic agent in treatment of

cancers that must obtain Asn from the blood, particularly acute lymphoblastic
leukemia. This cells express very low level of Asparagine synthetase.
Precursor functions: Asp is a neurotransmitter and a precursor of the pyrimidine synthesis.
In some cases (urea synthesis, purine synthesis) Asp is the amino-donor. In these reactions
aspartate in converted to fumarate.

Asp fumarate
energy
X-succinate
N
O
O NH N
-O NH2
Keto- or -O
N
carbonyl N
group O
N N
SUCCINYL-COA PATHWAY
General characteristics:
Degradation of essential amino acids,
mostly glucogenic features, but lipid-like degradations

Met Thr

Val -ketobutyrate Ile

propionyl-CoA (Ac-CoA)

succinyl-CoA
Common reactions:

Propionyl-CoA ------> succinyl-CoA. This it is also part of the degradation of

odd-numbered fatty acids. This pathway occurs in mitochondrion.

CO2 propionyl-CoA CH3

carboxylase
-
CH3 CH2 C CoA OOC CH C CoA
O biotin O
propionyl-CoA ATP ATP + Pi D-methylmalonyl-CoA

methylmalonyl-CoA
methylmalonyl-CoA racemase
mutase
- -
OOC CH2 CH2 C CoA OOC CH C CoA
O deoxyadenosyl-B12 CH3 O
succinyl-CoA L-methylmalonyl-CoA
ENZYME DEFICIENCIES OF THE PROPIONYL-COA
PATHWAY AND RELATED VITAMIN DEFICIENCIES
Enzyme deficiency of any of the enzymes of the propionyl CoA pathway causes acidosis
(e.g. propionic acidemia, methylmalonic acidemia)

Vitamin B12 deficiency has similar consequences to the methylmalonyl CoA mutase
deficiency. However, it also has an effect on the C1 metabolism.

-
OOC CH C CoA deoxyadenosyl-B12 -
OOC CH2 CH2 C CoA
CH3 O O
methylmalonyl-CoA
L-methylmalonyl-CoA mutase succinyl-CoA

Met synthetase
FH4
methyl-B12 homoCys

Met
methyl-FH4 B12
 -

DEGRADATION OF VALINE
-
COO COO
+ +
H3N CH H3N CH

AND ISOLEUCINE CH
CH2 CH3
CH
CH3 CH3
Valine and isoleucine degradation occurs in a similar way, CH3 1 COO
-
Val 1 COO
-

and their pathways also show similarities to the Ile -KG C O -KG C O
Glu CH Glu CH
degradation of Leu (at the AcAcCoA pathway). CH2 CH3
CH3 CH3
Val is glucogenic, Ile is both glucogenic and ketogenic, S-CoA CH3 S-CoA 2
2 NAD+
NAD+
Leu is purely ketogenic. C O
CoA-SH C O CoA-SH
CH NADH+H+
NADH+H CH
+
1 Transaminases: high activity in muscle CH2 CH3 CO2 CH3 CH3
CO2
CH3 3 3
2 Branched chain α-keto acid dehydrogenase enzyme FAD
FADH2
S-CoA FAD S-CoA
FADH2 C O
complex: high activity in liver mitochondria. C O
C
Further steps of degradation: mainly β-oxidation C
CH CH3
CH CH3 2
steps: FAD FADH2 CH3
H H2O S-CoA S-CoA 4 H2O
OH
C C C
C O 4 H2O C O
CH
C 3 4 HO CH CH3
CH
* O
C C
H H HO CH2 CH3
CH3 C O
CoA-SH CH
(trans) L-hydroxy- 5
NAD NAD+ HO CH2 CH3
NADH+H+ S-CoA 5 NAD+
O
5 C O
C O NADH+H+
NADH + H+ CH
CH
O C CH3
O O CH CH3
CH3
C
CoA-SH 6 *
C S-CoA CO2
CH3 C S-CoA
H
C O
-ketoacyl- O
CH2
* * Unique steps of the pathways CH3
ENZMYE DEFICIENCIES OF VALINE AND
ISOLEUCINE DEGRADATION
Transaminase deficiency: hipervalinemia,
hiperleucine - isoleucinemia.

Deficiency of the branched-chain α-ketoacid dehydrogenase enzyme complex:

Maple Syrup Urine Disease. The most common enzyme deficiency of this pathway.
Severe acidosis, mental retardation.
Thiamine administration may be helpful in some cases.

MENNO SIMONS (1496 – 1561)

ACETOACETYL-COA PATHWAY
General characteristics: essential and conditionally essential amino acids
ketogenic characters:
Leu, Lys are purely ketogenic
similarities to fatty acid degradation

Lys Trp (Ala) (pyruvate)

Phe
(fumarate)
-aminoadipate
Tyr

Acetoacetyl-CoA Leu

Acetyl-CoA
 DEGRADATION OF LEUCINE

Degradation of leucine occurs in the mitochondrion.

NAD+ NADH+H+
+ -KG Glu CoA-SH CO2
NH3 O O
- -
CH3 CH CH2 CH COO 1 CH3 CH CH2 C COO 2 CH3 CH CH2 C S-CoA
CH3 CH3 CH3 isovaleryl-CoA
FAD
leucine -ketoisocaproate
3
FADH2

ADP + Pi CO2
O H2O O ATP O
OH
-
CH2 C S-CoA 4 -
OOC CH2 C CH C S-CoA * CH3 C CH C S-CoA
OOC CH2 C
CH3 CH3 biotin CH3
-hydroxy--methylglutaryl-CoA -methylglutaconyl-CoA -methylcrotonyl-CoA
(HMG-CoA)
*

O O
-
OOC CH2 C CH3 + CH3 C S-CoA
acetoacetate acetyl-CoA
ENZYME DEFICIENCIES OF LEUCINE
DEGRADATION
Transaminase deficiency: hiperleucine - isoleucinemia.

Deficiency of the branched-chain α-ketoacid dehydrogenase enzyme complex:

Maple Syrup Urine Disease.

Isovaleryl-CoA dehydrogenase deficiency: Isovaleric acidemia: Severe acidosis,

mental retardation. „Sweaty feet syndrome”

Carboxylase deficiency: 3-Methylcrotonylglycinuria,

The patient presents an odor like male cat urine.
OUTLINE OF THE Ile Val
1
Leu

DEGRADATION 1 1
-ketoacids
transamination with -KG
oxidative decarboxylation: elágazó láncú
CoA-SH NAD+ -ketoacid dehydrogenase enzyme complex
OF VAL, ILE TPP, lipoamide, FAD 2 (BCKDC) TPP, NAD+, FAD, lipoamide and CoA-SH
cofactors
NADH + H+
AND LEU CO2
acyl-CoA derivatives
FAD FAD
3 3 3 dehydrogenation with FAD
FADH2 FADH2
-unsaturated acyl-CoA derivatives
ATP, CO2
* biotin
ADP + Pi
4 H2O 4 carboxylated derivative
H2O 4

-hydroxyacyl CoA HMG-CoA

*
CoA-SH Ac-CoA
-hydroxyacid acetoacetate
NAD+
5 5
NADH + H+ AcAc-CoA

-ketoacyl CoA semialdehyde

CoA-SH
CoA-SH * NAD+
6
CO2
Ac-CoA NADH + H+
propionyl CoA
DEGRADATION OF LYSINE
There is no transaminase for Lys. Degradation mainly occurs through saccharopine.
The result of the reaction is the same as would be of an ε-transamination.
- - -
COO COO COO
H +
CH2 NH2 O C H2O CH2 NH CH H2O C O H3N CH
+
CH2 CH2 NADPH+H NADP+ CH2 CH NAD+ NADH+H CH2
+ CH2
2
CH2 + CH2 CH2 CH2 CH2 + CH2
saccharopine saccharopine
CH2 - CH2 - CH -
COO dehydrogenase-I. COO dehydrogenase-II. 2 +
COO
+ +
HC NH3 HC NH3 HC NH3 glutamate
- -ketoglutarate - -
COO COO COO
-aminoadipate semialdehyde
lysine saccharopine NAD+
H2O

NADH+H+
-
COO

CH2
CH2
CH2
+
HC NH3
-
COO
-aminoadipate
FATE OF α-AMINOADIPATE
It is a monoamino-dicarboxylate, a higher homolog of Glu. the α-ketoacid product
of transamination (α-ketoadipate) is degraded in a similar way to the degradation
of branched-chain α-ketoacids derived from Val, Ile and Leu.

-
COO 1. transamination (PLP)
2. oxidative decarboxylation (enzyme complex)
3. FAD-dependent dehydrogenation
CH2 4. decarboxylation (PLP)
OH 5. water addition
CH2 6. NAD-dependent dehydrogenation
H product: acetoacetyl CoA
CH2
+
HC NH3
-
COO
α-aminoadipate
MINOR PATHWAY: WITH L AMINO ACID OXIDASE:
ELIMINATION OF THE α AMINO GROUP

FMN FMNH2 H2O

H2O

Lysine α-keto-ε-amino-
a cyclic compound α-aminoadipate
caproate
L-amino acid semialdehyde
oxidase
(peroxysomes)
-
N COO

(piperidine carboxylate)

double bond rearrangement with

oxido-reduction opening the ring
on the other side
PRECURSOR FUNCTIONS OF LYSINE:
SYNTHESIS OF CARNITHINE
Carnithine is used for fatty acid transport through the mitochondrial membrane (see lipid metabolism). The ε-
amino group of some Lysyl residues in proteins are mono- di- or trimethylated, in a SAM-dependent pathway
(for unknown reasons). This is a reversible covalent modification: the methyl groups can be removed.
CH3 CH3 CH3
+
CH3 N CH3 + +
CH3 N CH3 CH3 N CH3
CH2 succinate
O2 -KG CH2 glycine CH2
CH2 CO2
Fe2+ CH2 FH4 CH2
CH2 CH2 CH2
vitamin C serine hydroxymethyl
CH2 HC OH transferase
+
HC O
HC NH3 +
HC NH3
COO
-
COO
- -butirobetaine aldehyde
NAD+
-N-Trimethyllysine
-hydroxy--N-trimethyllysine NADH+H+

CH3 CH3
+ +
CH3 N CH3 CO2 succinate CH3 N CH3
-KG
CH2 O2 CH2
Fe2+
HC OH vitamin C CH2
CH2 CH2
- -
COO COO
L-Carnithine -butirobetaine
FA TRANSPORT TO THE MITOCHONDRIA
PROLINE (AND LYSINE) SIDE CHAIN
HYDROXYLATIONS IN COLLAGEN MATURATION
REACTIONS CATALYZED BY SERINE
HYDROXYMETHYL TRANSFERASE:
FH4 methylene-FH4
PLP
Ser Gly

FH4 FH4 + acetaldehyde

PLP
Thr Gly

FH4 FH4 + -butirobetaine aldehyde

PLP
-hydroxy--N-trimetyl- Gly
lysine
KIDNEY AND LIVER PROVIDES CARNITHINE
FOR THE OTHER TISSUES
DEGRADATION OF TRYPTOPHAN

The degradation of Trp does not resemble to that of any other amino acids.

O
O
- O
O2 O formate
-
H2O
O NH3
+ -
O O
+ N
NH3 +
N triptophan formamidase O NH3
dioxygenase HC NH2
tryptophan O
kynurenine
formylkynurenine
O2
NADPH + H+
kynurenine
FAD hydroxylase
NADP+ H2O
Ala H2 O O
-
COO PLP
-
O
kynureninase
+
NH2 O NH3
HO NH2
OH
3-hydroxyanthranilate 3-OH-kynurenine
FURTHER DEGRADATION OF
3-HYDROXYANTHRANILATE:

-
COO
1. dioxygenation forming semialdehyde
2. decarboxylation
3. aldehyde oxydation (NAD+ dependent)
O
O
NH2 4. reduction of the double bonds yielding
α-aminoadipate
OH

3-hydroxyanthranilate
 PRECURSOR FUNCTONS OF TRIPTOPHANE:
SYNTHESIS OF NMN

double bond
isomerisation,
O2 PRPP PPi
ring closure

3-hydroxyanthranilate semialdehide quinolinate desamido-NMN

Trp dioxygenase O
O O O CO2
O
O
O O O
O P O N
O
NH2 O O NH2 N O
O O
OH O

HO OH

Trp is also a precursor of serotonin and melatonin.

BH 4 O 2 H 2O qBH 2 CO 2
PLP
Trp OH-Trp OH-tryptamine N-acetylserotonin melatonin
tryptophan (serotonin) hydroxyindole-
hydroxylase N-acetyltransferase
O-methyltransferase

Serotonin is a potent vasoconstrictor and stimulator of smooth muscle contraction.

Melatonin release from the pineal is an example of biorythm.
 OH
DEGRADATION OF PHE, TYR, O
O
ENZYME DEFICIENCIES, NH +
O
O
3
PRECURSOR phenylalanine
O NADH + H + OH
FUNCTIONS:
2
phenylalanine homogentsate
hydroxylase tetrahidrobiopterine
O2
NAD+ H2O homogentisate
Phenylalanine and tyrosine O dioxygenase
degradation predominantly
occurs in liver. O
NH3+ O
HO O

The major pathway starts with tyrosine O

phenylalanine hydroxylase tyrosine KG

aminotransferase O
O O
Glu maleylacetoacetate
O
maleylacetoacetate
isomerase GSH
O
O
HO
O O
O2 O
p-hydroxyphenylpyruvate O
dioxygenase O
CO2
O
OH fumarylacetoacetate

There is a minor pathway of O fumarylacetoacetase

the phenylalanine degradation O
O
starting with transaminase. OH O O
The capacity of this enzyme is homogentisate O O
low.
O O
fumarate
acetoacetate
METABOLISM OF TYROSINE
ENZYME DEFICIENCIES OF THE
PHE DEGRADATION PATHWAY
Phenylalanine hydroxylase deficiency: classical phenylketonuria (PKU). Most common
desease of the amino acid metabolism. Autosomal recessive. Mental retardation,
light skin color. Therapy: synthetic diet low in phenylalanine.
Dihydrobiopterine reductase deficiency: only 3% of PKU. Tyr and Trp hydroxylation is
also deficient, therefore central nervous functions are more seriously affected.
Therapy involves 5-hydroxytryptophan and DOPA administration.
Maternal PKU: Even if the neonate is a heterozygote, high Phe concentration in the mother's
blood causes severe mental retardation and heart problems of the fetus.
Enyzmes of the BH4 synthesis: In very rare cases the synthesis of the cofactor from GTP
may also be affected due to the deficiency of one of the enzymes catalyzing the process.

Tyrosine aminotransferase deficiency: tyrosinemia type 2. Eye, skin lesions, usually mental
retardation.

Fumaryloacetoacetase deficiency: tyrosinemia type 1. liver and kidney failure, problems

affecting the nervous system, yellowing of the skin and whites of the eyes (jaundice),
cabbagelike odour, and increased tendency to bleed (particularly nosebleeds).

Homogentisate dioxigenase deficiency: alcaptonuria. It was the first identified inborn error
of metabolism. Autosomal recessive. Ochronosis, arthritis. Relatively mild disease.
SMELL AS A DIAGNOSTIC MARKER

Disorder Urine Odor

Phenylketonuria (PKU) "musty" or "mousy" smell

Cystinuria "sulfur" smell

Isovaleric acidemia, "sweaty feet" smell

3-Methylcrotonylglycinuria „male cat urine” smell

Methionine malabsorption "cabbage" smell

Tyrosinemia "cabbage" or "rancid butter." smell

Methylmalonic acidemia acidic odor

MSUD "curry," "maple syrup",

"burnt sugar", or "fenugreek."
PRECURSOR FUNCTIONS OF PHE AND TYR
Synthesis of neurotransmitters: synthesis of catecholamines (3-4-dihydroxy derivatives of
phenylalanine):
O
OH
O
NH3+
HO
tyrosine NH2-CH3
HO
NADH + H+ OH
tyrosine O 2
epinephrine
hydroxylase tetrahidrobiopterine SAH
phenyletanolamine
NAD+ H2O N-methyltransferase
SAM
O OH
O
NH3+
HO NH3+
OH HO
OH
3,4,-dihydroxyphenylalanine (DOPA) H2O norepinephrine
CO2 O2
DOPA
decarboxylase dopamine  - hydroxylase

NH3+
HO
OH
dopamine
Parkinson’s disease: Due to degeneration and death of cells in substantia nigra and locus
coeruleus the amount of dopamine released decreases as a function of the number of surviving
cells. Treatment: administration of dopa with analogues that inhibit dopa decarboxylase and are
unable to cross the blood-brain barrier.
MELANINE SYNTHESIS
O O O

O O O
NH3+ melanine
NH3+ NH3+
HO tyrosinase H O tyrosinase O
OH O
tyrosine dopa dopa quinone

Tyrosinase deficiency: classical albinism. Impaired melanine formation,

sensitivity to sunlight.
MAJOR POINTS:
• Essential/nonessential, glucogenic/ketogenic nature of amino acids
• Features of transaminases and glutamate dehydrogenase, role of trans-
deamination
• How can PLP be a cofactor for various enzymes?
• Formation of ammonia in the human body
• Major features of the amino acid metabolism of tissues, N-transport between
the tissues
• CPS-I reaction and the urea cycle, related enzyme deficiencies
• C1 transfer, transmethylation, related enzyme and vitamin deficiencies
• Aromatic amino acid oxidase enzyme family, types of PKU
• Role of dioxigenase reactions in amino acid metabolism. Alkaptonuria
• General features of the pyruvate pathway
• Transsulfuration
• General features of the alpha-ketoglutarate pathway,
• General features of the succinyl-CoA pathway
• Vitamin B12-dependent reactions in the human body, consequences of
the vitamin B12 deficiency
• General features of the acetoacetyl-CoA pathway
• Degradation of the branched chain amino acids (Val, Leu, Ile)
• Major outline of Trp degradation
• Steps of Phe degradation
• Inborn errors of enzymes of aminoacid metabolism (PKU, MSUD...)
SHORT QUESTIONS (1):
1. What are the aspects of the essential feature of an amino acid?
2. Examples of circumstances causing positive or negative N balance
3. What type of amino acid transporters are known?
4. What are the main features of transaminases?
5. What are the main features of glutamate dehydrogenases?
6. What is transdeamination?
7. What is the meaning of stereoelectronic control in PLP-dependent
reactions?
8. What is the molecular basis of ammonia toxicity?
9. What are the enzyme activities required for mitochondrial
carbamoyl-phosphate synthesis?
10. List the enzymes of the urea cycle!
11. What are the special methyl donors in the body?
12. What enzyme deficiencies may lead to homocystinemia?
(at least two examples)
13. List SAM-utilizing reactions! (at least three examples)
14. What are the common consequences of folic acid and vitamin B12
deficiencies?
15. What is the methyl-tetrahydrofolate trap?
16. What reactions are affected by dyhydrofolate reductase deficiency?
SHORT QUESTIONS (2):
17. What type of reactions are involved in the degradation of branched-chain
amino acids (Leu, Ile, Val)?
18. What enzyme deficiencies may lead to hyperphenylalaninemias?
19. What kind of genetic and none-genetic causes may lead to
hyperammonemia?
20. What is the molecular reason of alkaptonuria? What are the symptoms?
21. What determines the glucogenic/ketogenic nature of an amino acid?
22. What are the molecular consequences of vitamin B12 deficiency?
23. What is the reason why the human body cannot produce large amount of
arginine?
24. How is polyamine synthesis regulated?
25. What is the main reason why DFMO turned out to be useless as
antitumor drug?
26. What happens in the trans-sulfuration reaction?
27. What are the C1 sources in the body?
28. What is the cofactor of serine hydroxymethyl-transferase and what kind of
reactions does it participate?
29. What is the active sulfate in the body, how is it produced?
30. Examples for metabolic acidosis in the enzyme deficiencies of amino acid
metabolism (at least two examples)
31. List the reactions affected by the dihidrobiopterin reductase deficiency?
32. List enzymes utilizing PLP as cofactor (at least four examples)