Amino acid metabolism II

Metabolism of individual amino acids

Biochemistry I
Lecture 7 2008 (J.S.)
The degradation of amino acids usually begins with deamination.
However, transamination or oxidative deamination is not the first
reaction in catabolism of eight amino acids:
Serine and threonine are deaminated by dehydration, and
histidine undergoes deamination by desaturation
(both reactions were mentioned previously).
The five remaining amino acids are deaminated later on, after partial
transformation:
Arginine – deamination occurs after transfomation to ornithin,
lysine – transamination follows the transformation to α-aminoadipate,
methionine – deamination of homoserine,
proline – deamination after conversion to glutamate,
tryptophan – after its transformation to kynurenine, alanine is
released.

2
Each carbon skeleton of deaminated amino acids follows
a unique metabolic pathway to compounds , which can be
completely oxidized by way of the citrate cycle to CO2 and water.

In spite of this common fate, amino acids are classified as

glucogenic and ketogenic according to the type of their
intermediate metabolites.

The glucogenic amino acids give rise to pyruvate or some of

the intermediate of the citrate cycle, which can serve as
substrates for gluconeogenesis.
The ketogenic amino acids give rise to acetoacetate or
acetyl-CoA (from which acetoacetate can be synthesized)
that cannot be transformed to glucose.

3
Glucogenic and ketogenic amino acids

4
Irreversible conversions in the metabolism of amino acids
show which proteinogenic amino acids are essential:

5
Nonessential amino acids Essential amino acids:

Glycine Threonine
Alanine
Serine Methionine
Cysteine Lysine
Aspartate Valine
Asparagine Leucine
Glutamate Isoleucine
Glutamine
Histidine
Proline
Arginine Phenylanine
Tyrosine Tryptophan

6
The metabolism of amino acids will be described
in the following sequence:

1 The most simple AA that give pyruvate – Ala, Ser, Gly, Thr
2 Amino acids containing sulfur – Met, Cys
3 Sources of one-carbon units and use of those units in syntheses
4 Aspartic acid
5 Glutamic acid and its relation to Arg, Pro, His
6 Branched-chain amino acids – Val, Ile, Leu
7 Lysine
8 Aromatic amino acids – Phe, Tyr, and Trp

7
 1 Amino acids that are converted to pyruvate:

Alanine - by transamination.
Serine - by deamination catalyzed of dehydratase (hydrolyase).
Glycine - by accepting one-carbon group gives serine.
Threonine - by splitting gives glycine that may give serine.

Cysteine also gives pyruvate by deamination and desulfuration

(see "Amino acids containing sulfur"), as well as
tryptophan that after transformation to kynurenin releases alanine
(see "Aromatic amino acids").

8
 Alanine is nonessential and glucogenic;
it undergoes transamination to pyruvate readily:
H3C–CH–COOH alanine aminotransferase (ALT)
NH2
H3C–CH–COOH H3C–C–COOH
NH2 Glu O pyruvate
2-oxoglutarate

Concentrations of alanine in blood plasma are 300 – 400 μmol/l (the

second highest next to glutamine). Alanine is released from muscle
tissue and serves both as the vehicle for NH3 transport from muscle
to liver and a substrate for liver gluconeogenesis. This bidirectional
transport is called the alanine cycle (or glucose-alanine cycle).
Blood
Muscle Liver
glucose glucose
glycolysis gluconeogenesis
pyruvate pyruvate

alanine alanine

9
 Serine
CH2–CH–COOH
OH NH2 is nonessential and glucogenic;
– nonessential – synthesis of the carbon skeleton from 3-phosphoglycerate
– glucogenic – direct deamination by serine dehydratase to pyruvate

Serine does not take part in transamination,

but it is directly deaminated by dehydration:

H2O
NH3
CH2–CH–COOH CH2=C–COOH CH3–C–COOH CH3–C–COOH

=
OH NH2 NH2 NH O
H2O enamine imine
serine pyruvate

10
Serine is a substantial source of one-carbon groups:
its -CH2-OH group is readily transferred to tetrahydrofolate
(coenzyme of C1-group transferase), the product is glycine that
is able to yield the second C1-group.
The reaction is reversible, but the synthesis of serine from glycine
and a C1-group is not an advantage.
CH2–CH–COOH CH2–COOH
+ H4folate + CH2OH–H4folate
OH NH2 NH2
serine glycine hydroxymethyl-H4folate

Decarboxylation of serine results in ethanolamine

(a constituent of phospholipids) that gives choline
by methylation. + CH3
HO–CH2–CH2–NH2 HO–CH2–CH2–N– CH3
ethanolamine CH3
choline
11
 Demands for serine in the body are great – both one-carbon
groups and substrates for the synthesis of complex lipids have to
be supplied.
Therefore, the synthesis of carbon skeleton from glucose is of
great significance:
2-oxoglutarate
NAD
+
Glu
COOH COOH COOH
glucose CH–OH O– C=O O– CH–NH2 serine
O–
CH2–O–P=O CH2–O–P=O CH2–O–P=O
O– O– O–
3-phosphoglycerate 3-P-hydroxypyruvate 3-phosphoserine

12
 Utilization of serine:
glucose
phosphatidyloserines
3-phosphoglycerate

phosphohydroxypyruvate
Glu
phosphatidylethanolamines
phosphoserine
ethanolamine
glycine serine phosphatidylcholines
choline
CH2-OH–H4folate
acetylcholine
NH3
palmitoyl-CoA
pyruvate

sphingosine sphingomyelins

13
Glycine
CH2–COOH is nonessential and glucogenic;
NH2 – nonessential – originates from serine or from CO2, NH3, and C1-group
– glycogenic (weakly) – may accept C1-group and give serine
Reversible reaction
glycine + CH2OH–H4folate serine + H4folate
(described as an important source of C1-groups) is not a useful
way of glycine catabolism, because it consumpts one C1-group.
Transamination of glycine with pyruvate
glycine + pyruvate glyoxylate + alanine
as well as oxidative deamination of glycine
glycine + FAD glyoxylate + FADH2
are possible, although limited; the enzymes catalyzing those reactions
have sufficient activity only in peroxisomes. It is worth mentioning that
glyoxylate formed in those minor pathways gives small amounts of
unwanted oxalate. High production of oxalate is dangerous.
14
 The major pathway of glycine catabolism is
oxidative cleavage of glycine in mitochondria:

CH2–COOH
+ H4folate CO2 + NH3 + N5,N10-methylene-H4folate
NH2
glycine

The reaction is reversible and catalyzed by glycine synthase and

controlled by respiration and energetic charge of the cell. For
the synthesis of glycine, 3 molecules of ATP are lost.

Molecule of glycine is the substrate required for the syntheses

of several very important compounds, e.g.
purine bases of nucleic acids, porphyrins of haemoproteins,
phosphocreatine of skeletal muscles (phosphagen), and
tripeptide glutathione (intracellular antioxidant).

15
 (oxaluria)
Utilization of glycine:

glyoxylate

Ala Synthesis of
creatine
pyruvate
purine
serine glycine porphyrin
glutathione
hydroxymethyl-H4folate
methylene-H4folate glycine conjugates
CO2 NH3 of bile acids,
of aromatic acids
Gly oxidative splitting / Gly synthesis
(mitochondria) (hippuric acids)

16
 Threonine is essential and both glucogenic and ketogenic
CH3–CH2–CH–COOH It does not undergo transamination

OH NH2 – glucogenic – gives glycine by splitting

or succinyl-CoA (by dehydratation and
and oxid. decarboxylation to propionyl-CoA)
– ketogenic – by splitting to glycine gives acetyl-CoA

Splitting of threonine to glycine

CH3–CH2–CH–COOH Ser CH2OH transferase CH2–COOH CH2OH-H4folate

OH NH2 NH2 serine

glycine
Thr dehydrogenase CH3–CH=O
acetaldehyde
specific lyase oxid. pyruvate
2-amino-3-oxobutyrate
oxid.

CH3-CO–S-CoA
acetyl-CoA

17
 An alternative pathway is
the direct deamination of threonine by dehydration:

H2O NH3
CH3–CH2–CH–COOH CH3–CH2–C–COOH

=
[ enamine imine ]
OH NH2 O
H2O 2-oxobutyrate

(slow) oxidative
decarboxylation

CH3-CH2-CO–S-CoA
propionyl-CoA

carboxylation to methylmalonyl-CoA
rearrangement (B12 coenzyme)

HOOC-CH2-CH2-CO–S-CoA
sukcinyl-CoA

18
 2 Sulfur containing amino acids
Methionine CH2–CH2–CH–COOH is essential and glucogenic
CH3–S NH2 – glucogenic (it yields succinyl-CoA)

Methionine is a common methyl donor in the cell:

PPi + Pi CH3
CH2–S Rib-Ade
ATP
CH2
CH-NH2 substrates
CH2–S–CH3
CH2 COOH
methylated substrates
CH-NH2 S-adenosylmethionine
COOH
CH2–S
methionine Rib-Ade
CH2
CH2–SH CH-NH2
H4folate
CH2 COOH
CH-NH2 S-adenosylhomocysteine
CH3-H4folate
remethylation (B12 coenzyme) COOH Ado

homocysteine 19
Activated methionine
NH2
S-adenosylmethionine (S-AM)
N
N
CH3
+ N
HOOC-CH-CH2-CH2–S N
CH2
NH2 O
is the methyl donor. The methyl
group may be transferred from
a sulfonium ion to various acceptors. OH OH

Examples:
synthesis of choline from phosphatidylethanolamine,
synthesis of creatine (by methylation of guanidinoacetate),
methylation of noradrenaline to adrenaline,
inactivation of catecholamines by catechol-O-methyl transferase,
methylation of histones, etc.

20
Catabolism of methionine
– demethylation to homocysteine
methionine – transsulfuration with serine to homoserine and cysteine
– conversion to 2-oxobutyrate (homoserine deaminase),
S-AM propionyl-CoA, and succinyl-CoA.
–CH3 + Ado

– H2O

serine
homocysteine
cysteine

cystathionine succinyl-CoA
+ H2O
(coenz. B12)

methylmalonyl-CoA
CoA-SH CO2

CO2 (biotin)
NAD+ NADH + H+
NH3 propionyl-CoA
homoserine 2-oxobutyrate 21
Homocysteine is an important intermediate in metabolism of methionine;
it is readily transformed, either remethylated to methionine
(the reaction requires tetrahydrofolate and cobalamin)
or decomposed to homoserine by transsulfuration with serine,
(vitamin B6 dependent).

If those mechanisms are not sufficient and the concentration of homocysteine in

biological fluids increases, injury of endothelial cells by homocysteine (e.g., high
production of reactive oxygen species, lipoperoxidation) and decreased vitality
of blood platelets may appear.

At present, high concentration of homocysteine in blood plasma is

included among other biochemical markers of cardiovascular diseases –
as a risk factor for atherosclerosis that is quite independent on the
concentration of cholesterol.

22
 Cysteine is nonessential and glucogenic
– nonessential – synthesis from serine
CH2–CH–COOH (methionine supplies the sulfur atom)
SH NH2 – glucogenic – cysteine is converted into pyruvate
(sulfur atom is released as SO32–, HS–, or SCN–)

The major catabolic pathway is the direct oxidation of SH-group:

CH2–SH
CH-NH2 O H+
SO42–
H+O -S=O sulfate
COOH ll
O
oxidation of SH-group
cysteine (mitochondrial dioxygenase)
sulfite oxidase

O2 + NADPH + H+ O H+ SO32–
H+O -S sulfite
ll
O
O O
ll ll
S-O H+ S-O H+ CH3
CH2 2-OG Glu CH2 C=O
CH–NH2 C=O
transamination COOH
COOH COOH
cysteine sulfinate pyruvate
3-sulfinylpyruvate 23
 Oxidation of S–II to SIV or SVI (sulfinate, sulfite, sulfate) is
a proton-producing process, nonvolatile acids are formed from
non-ionized groups. The catabolism of sulfur-containing amino acids
slightly acidifies the body.

An alternative catabolic pathway of cysteine is transamination :

CH2–SH 2-oxoglutarate CH2–SH CH3

Glu
CH-NH2 C=O desulfuration C=O
COOH transamination COOH COOH
cysteine 3-sulfanylpyruvate pyruvate
HS– H+

Hydrogen sulfide HS– ion is mostly oxidized to sulfite SO32– or, if cyanide ion CN–
is present (e.g. tobacco smokers), hydrogen sulfide gives thiocyanate SCN–.

Sulfite anion is oxidized to sulfate anion, which is either excreted into

the urine (approx. 20 – 30 mmol/d) or utilized for sulfations after activation:
mitochondrial sulfite oxidase excretion
SO32– SO42–
(molybdopterin, cyt b5)
3´-phosphoadenosyl
sulfite sulfate (ATP) 5´-phosphosulfate, PAPS
24
 3‘-Phosphoadenosyl-5‘-phosphosulfate (PAPS)
NH2
is the mixed anhydride of sulfuric and N
N
phosphoric acid called "active sulfate"; O
it serves as the sulfate donor in forming
O
N
5' N
of sulfate esters (or N-sulfates). O–S–O–P–O– CH2 O
O O
3'
O OH

O–P=O
O
Examples of sulfations by means of PAPS:
synthesis of proteoglycans (sulfation of glycosaminoglycans),
sulfation of saccharidic components in glycolipids and glycoproteins,
formation of sulfate esters in inactivation of steroid hormones, catecholamines,
and in the phase II of biotransformation of phenols.

25
 Utilization of methionine and cysteine

serine – 2H cystine
methionine
decarbox.
cysteine 2-aminoethanethiol
(cysteamine, constituent of coenzyme A)
SAM homoserine
methylations
glutathione
(γ-glutamyl-cysteinyl-glycine)
succinyl-CoA
3-cysteine
sulfinate
mercapturic acids
(N-acetyl-S-arylcysteines)

decarbox.
SO4 2–
SO 3
2–

pyruvate
PAPS hypotaurine taurine
sulfate esters conjugation with
bile acids
of sugars and phenols
26
 Glutathione (GSH, γ-glutamyl-cysteinyl-glycine)
γ
is a tripeptide with a free sulfanyl group,
CO–NH–CH–CO–NH–CH2–COOH required to maintain the normal reduced
CH2 CH2–SH state in the cell:
CH2 – 2H
α (reduced form) 2 G-SH G-S–S-G
CH–NH2 + 2H

COOH Functions:
1 Reduced G-SH confronts oxidative stress,
it reduces peroxides (lipid hydroperoxides and hydrogen peroxide) in the reaction
catalyzed by a selenoprotein glutathione peroxidase, and (non-enzymatically)
methaemoglobin (FeIII, hemiglobin) to haemoglobin (FeII) and disulfides to thiols:

L-OOH + 2 G-SH L-OH + G-S–S-G + H 2O

R-S–S-R + 2 G-SH 2 R-SH + G-S–S-G
reduced G-SH can be regenerated by glutathione reductase and NADPH + H+.
2 Conjugation to lipophilic compounds (detoxification of reactive electrophiles).
3 Transport of amino acids into cells with concomitant attachment of γ-glutamyl
(group translocation, γ-glutamyl cycle). 27
3 Sources of one-carbon groups and
utilization of those groups in syntheses
One-carbon groups are transferred by tetrahydrofolate (H4folate, FH4,
tetrahydropteroylglutamate).
Mammals can synthesize the pteridine ring, but they are unable to
conjugate it to the other two units. They obtain folate from diets or from
microorganisms in their intestinal tracts.

Sites for bonding of one-carbon units

COOH
OH H
N CH2-NH– –CO–NH-CH-CH2-CH2-COOH
N 5 10

H2N
N N
H
glutamate
tetrahydropteroic acid (1 – 5 residues)

28
The one-carbon groups transferred by H4folate exist in three oxidation states:

Example:
CH2 COOH
OH
N CH2–N– –CO–NH-CH-CH2-CH2-COOH
N 5 10

H2N
N N
H N5,N10-methylene FH4

(The fully oxidized one-carbon group is CO2, but CO2 is transferred by biotin,
not by H4folate.)
29
 methylations

homocysteine S-AM

methionine

N5-methyl FH4 thymine

glycine, serine nucleotides
NADH

N5,N10-methylene FH4 purine

nucleotides
NADPH
NH3 H2O
N5-formimino FH4 N5,N10-methenyl FH4 N10-formyl FH4

histidine tryptophan
30
4 Aspartic acid and asparagine
Aspartate is nonessential and glucogenic
– it gives oxaloacetate by transamination
HOOC–CH2–CH–COOH
NH2 Asparagine is the amide of aspartate
H2N-CO–CH2–CH–COOH
NH2

asparagine 2-oxoglutarate glutamate

asparaginase
H2O NH3

AST

glutamate glutamine aspartate oxaloacetate

+ AMP + 2 Pi + ATP
Asn synthetase

31
Utilization of aspartate and asparagine

NH3 for the synthesis of

urea
malate fumarate AMP from IMP
purine
AST
oxaloacetate aspartate incorporated into
the skeleton of
Glu pyrimidine bases
NH3 Glu
CO2
β-alanine
asparagine
NH3 transport in CNS ?

32
5 Glutamic acid, glutamine, and the relationship
to proline, arginine, and histidine
Glutamate is nonessential and glucogenic
– it gives oxaloacetate readily by oxidative
HOOC–CH2–CH2–CH–COOH deamination or transamination
NH2
Glutamine is an amide of glutamate
H2N-CO–CH2–CH2–CH–COOH
NH2
glutamine
pyruvate alanine
glutaminase
H2O NH3

ALT

ADP + Pi NH4+ + ATP

glutamate 2-oxoglutarate
Gln synthetase
33
 Direct oxidative deamination of glutamate by dehydrogenation

The reaction is catalysed by the mitochondrial enzyme

glutamate dehydrogenase (GLD). It requires either NAD+ or NADP+ as coenzyme,
and its activity in mitochondria is high.

HOOC-CH-CH2-CH2-COOH HOOC–C–CH2-CH2-COOH
NH2 NH 2-Iminoglutarate
NAD(P)+ NAD(P)H + H+
glutamate H2O

NH3

HOOC–C–CH2-CH2-COOH
The equilibrium favours glutamate
O
synthesis, but it is pulled in the direction
od deamination by the continuous 2-oxoglutarate
removal of NH3/NH4+.
34
Decarboxylation of glutamate (very active in brain)

– CO2
γ
γ-aminobutyric acid (GABA)
glutamate
an inhibitory neurotransmitter in CNS

Reversible reduction of glutamate

proline

NADH+H+ NAD+
2 H+ H2O ornithine

glutamate glutamate 5-semialdehyde

– intermediate in the synthesis
and degradation of proline and arginine
35
Utilization of glutamate and glutamine

ornithine proline

histidine glutamate semialdehyde

CO2
γ-aminobutyrate
(inhibitory neurotransmitter)
transaminases glutamate
2-oxoglutarate glutamate
(excitatory neurotransmitter)
AA 2-oxoA
NH3 pteroate
NH3

cysteine folate
glutamine glycine
transport of NH3 to the liver and kidney
donor of NH3 for the syntheses of glutathione
carbamoyl phosphate
purine
amino sugars 36
Glutamate is widely used as a food additive to enhance flavour of
dishes, particularly in Chinese cookery in high amounts.
Excess in the diet (1 – 5 g of glutamate in one dose, e.g. in the form
of "Von-Ton“ soup) can cause unpleasant feelings in sensitive
persons – the Chinese restaurant syndrome.

37
 Arginine
CH2–CH2–CH2–CH–COOH is nonessential and glucogenic
H2 N NH NH2 – nonessential in adult man (required in
C the diet during the growth)
NH – degraded to 2-oxoglutarate

In the liver, arginine is hydrolyzed to ornithine and urea. Ornithine serves as

the substrate for ureosynthetic cycle:

arginase
+ H2O
+

urea
arginine ornithine
CO2
decarboxylation

putrescine (butan-1,4-diamine) for

the ureosynthetic cycle synthesis of polyamines
38
 After hydrolysis of arginine to ornithine,
ornithine is degraded by transamination of the 5-amino group to
glutamate 5-semialdehyde that gives glutamate and 2-oxoglutarate.

Nitroxide (nitrogen monoxide, NO) originates from arginine:

O2. NADPH O2. NADPH •N=O +

nitroxide
(a radical)
arginine Nω-hydroxyarginine citrulline

The reaction is a five-electron oxidation catalyzed by nitroxide synthase (NOS),

employing five redox cofactors (NADPH, FAD, FMN, cytochrome, H4biopterin).
There are three isoenzymes of NOS: endothelial NOS responsible for
vasodilation and inhibition of platelet aggregation, neuronal NOS modulation
events on synapses (both are Ca2+-dependent), and NOS in phagocytes (NO
gives bactericidal peroxynitrite ONOO–). 39
Synthesis of creatine
Arginine is the donor of amidino group for the synthesis of creatine:

S-AdoHcy
(kidney) S-AM
(liver)

creatine
N -methylguanidinoacetate
1

Creatine in skeletal muscles:

ATP ADP

creatine kinase

phosphocreatine
(muscle phosphagen)
H2O

slow non-enzymatic
dehydration creatinine
(excreted into the urine)
40
 Proline (pyrrolidine-2-carboxylic acid)
is nonessential and glucogenic
– nonessential – originates from glutamate
N COOH – glucogenic – it gives 2-oxoglutarate
H

H2O
NAD+ NADH+H+

1-pyrroline 5-carboxylate glutamate 5-semialdehyde

proline (cyclic aldimine)
NAD+ + H2O
NADH+H+
AA 2-oxoA

2-oxoglutarate glutamate

4-Hydroxyproline
occurs only in collagen, and is formed by posttranslational
hydroxylation of prolyl residues in procollagen polypeptide
chains. Similarly to proline, 4-hydroxyproline is degraded to
4-hydroxyglutamate, which is cleft to pyruvate and glyoxylate.
41
 Histidine is nonessential and glucogenic
– nonessential for adults (essential for children)
CH2–CH–COOH – glucogenic - it gives glutamate and 2-oxoglutarate
N NH2

N Histidine mostly does not undergo transamination,

H it is deaminated directly by elimination (desaturation):

H2O
NH3-lyase

NH3 urocanic acid

histidine 4-imidazolone-5-propionate
(urocanate)
H2O

AA 2-oxoA

2-oxoglutarate glutamate

HN=CH–FH4 FH4 N-formimino-glutamate

N5-formimino tetrahydrofolate (FIGLU)

42
Histamine is the product of histidine decarboxylation catalyzed
by specific histidine decarboxylase:

CO2
histidine histamine

Histamine is a biogenic amine stored within granules of basophils and mast cells
(more than 90 % body stores) and within synaptosomes of certain CNS neurons.
When released, histamine induces complex physiological and pathological effects,
including immunological reactions (symptoms of allergic conditions of the skin
and airways), gastric acid secretion, smooth muscle contractions (e.g.
bronchoconstriction), and profound vasodilatation. Histamine exerts its action via
at least four distinct histamine receptor subtypes.
Released histamine is metabolized by oxidation (to imidazolylacetic acid) or
methylation (to tele-N-methylhistamine and tele-N-methylimidazolylacetic acid).

Antihistaminics – drugs which antagonize the effects of histamine.

43
Amino acids metabolized to 2-oxoglutarate – relationships:

proline histidine
arginine
urea

1-pyrroline-5-carboxylate urocanate
ornithine

glutamate semialdehyde N-formiminoglutamate

glutamate 2-oxoglutarate
2-oxoA AA

glutamine
44
 6 Branched-chain amino acids
Leucine Isoleucine
Valine CH3
CH3 CH3
are all essential, their
CH3 CH3 CH CH2 CH3
final metabolites are different:
CH CH2 CH
valine is glucogenic,
CH–NH2 CH–NH2 CH–NH2 leucine is ketogenic,
COOH COOH COOH isoleucine both gluco- and ketogenic.
These amino acids are taken up from the blood predominantly by skeletal muscles and
their catabolism (transamination) begins there.
The three initial catabolic reactions are common to all three
branched-chain amino acids:
– transamination to corresponding 2-oxoacids,
– oxidative decarboxylation catalyzed by 2-oxoacid dehydrogenase
producing corresponding acyl-CoA thioesters, and
– the second dehydrogenation between carbons α and β catalyzed
by flavin dehydrogenase resulting in corresponding 2-alkenoyl-CoA thioesters:
45
The resulting 2-alkenoyl-CoAs after three initial reactions:

valine leucine isoleucine

The following reactions differ (expected addition of water, hydration, occurs

as the next reaction only in the case of valine and isoleucine).

Leucine is ketogenic. The alkenoyl-CoA is carboxylated (CO2 donor is

carboxy-biotin) and the product hydrated to HMG-CoA that splits to
free acetoacetate and acetyl CoA:.

H2O acetoacetate

biotin hydration acetyl-CoA

CO2-biotin
carboxylation
3-hydroxy-3-methylglutaryl-CoA
(HMG-CoA) 46
 Valine

succinyl-CoA
(B12)
CO2
carboxylation
CoA-SH CoA-SH CO2
hydration oxidation decarboxylation propionyl-CoA methylmalonyl-CoA

Isoleucine

succinyl-CoA
CoA-SH acetyl-CoA (B12)

carboxylation

hydration oxidation splitting CO2

propionyl-CoA 47 methylmalonyl-CoA
 Branched-chain amino acids – summary:

isoleucine leucine

valine

3-hydroxy-3-methylglutaryl-CoA
(HMG-CoA)

acetyl-CoA
propionyl-CoA propionyl-CoA
acetyl-CoA
succinyl-CoA succinyl-CoA acetoacetate

glucogenic ketogenic ketogenic

glucogenic

48
 7 Lysine is essential and ketogenic
– it gives acetoacetyl-CoA
CH2–CH2–CH2–CH2–CH–COOH
NH2 NH2

Lysine does not undergo transamination.

Primarily, ε-deamination occurs through the formation of saccharopine:

glutamate
ε + NAD+
NADPH+H+ H2O H2O NAD+

reduction oxidation

α 2-oxoglutarate

lysine saccharopine allysine 2-aminoadipate

Transamination of α-amino group in 2-aminoadipate follows:

49
 NAD+
2-oG Glu CoA-SH
CO2

CO2 FAD

glutaryl-CoA crotonoyl-CoA
2-aminoadipate 2-oxoadipate
H2O hydratation
NAD+
dehydrogenation

Lysyl side chains in collagen and elastin are

oxidatively ε-deaminated to allysyl side chains.
The aldehyde groups so formed react non-enzyma-
tically with each other, or with lysyl ε-NH2, and acetoacetyl-CoA
form covalent crosslinks
(pyridinoline type in collagen, isodesmosine in elastin).

50
8 Aromatic amino acids
phenylalanine, tyrosine, and tryptophan

All three amino acids are essential (though tyrosine is also formed
by hydroxylation of phenylalanine),
and both glucogenic and ketogenic,
- phenylalanine and tyrosine give fumarate and acetoacetate,
- tryptophan gives alanine and acetoacetyl-CoA.

51
Phenylalanine and tyrosine
Hydroxylation of phenylalanine to tyrosine
is catalyzed by a monooxygenase – phenylalanine hydroxylase,
for which the reducing coenzyme is tetrahydrobiopterin (BH4):

phenylalanine

O2
5,6,7,8-tetrahydrobiopterin NAD+

NADH + H+
tyrosine
H2O

q-7,8-dihydrobiopterin

Similarly, tyrosine is hydroxylated to DOPA by tyrosine 3-hydroxylase, and

tryptophan to 5-hydroxytryptophan by tryptophan 5-hydroxylase.
52
 hydroxylation transamination
(O2, BH4) 4-hydroxy-
2-oG Glu phenylpyruvate

O2 dioxygenase
phenylalanine tyrosine
CO2

homogentisate
(2,5-dihydroxy-
phenylacetate)

O2
1,2-dioxygenase

H2O
fumarate maleinylacetoacetate
isomerization
acetoacetate fumarylacetoacetate
53
Inborn metabolic disorders of phenylalanine catabolism

(O2, BH4) (O2, BH4)

phenylalanine tyrosine DOPA
2-oG
Glu ALBINISM
PHENYLKETONURIA,
hyperphenylalaninaemias 4-hydroxyphenylpyruvate
O2
HYPERTYROSINAEMIA II
CO2
homogentisate
(2,5-dihydroxyphenylacetate)
O2 ALKAPTONURIA

maleinylacetoacetate

fumarylacetoacetate
H2O HYPERTYROSINAEMIA I
fumarate + acetoacetate

54
Hyperphenylalaninaemia type I
(classic phenylketonuria, PKU)
Phe hydroxylase
is a defect in phenylalanine hydroxylase, (O2, BH4)
the ability to convert Phe to tyrosine is
considerably impaired. (tyrosine)
PKU have to be recognized through the
compulsory screening of newborn infants
phenylalanine
and treated by a low-phenylalanine diet
till the age of 8 – 10 years. transamination

The consequence of untreated PKU is

mental retardation (oligophrenia
phenylpyruvica). Besides high levels of phenylacetate
blood Phe, alternative catabolites are
produced and excreted in high amounts
(a "mousy" odour of the urine) : phenylpyruvate

o-hydroxyphenylacetate
Malignant hyperphenylalaninaemias type IV and V
BH4 (tetrahydrobiopterin) is lacking due to the defective dihydrobiopterin
biosynthesis from guanylate, or an ineffective reduction of BH2 to BH4.
55
Hypertyrosinaemias
occur in several forms. They may be caused by a deficit of enzymes which catalyze
either the transamination of tyrosine, or oxidation of p-hydroxyphenylpyruvate and
hydrolysis of fumarylacetoacetate. A low-tyrosine diet may be very useful.
Plasma levels of tyrosine are elevated, and large amounts of tyrosine,
p-hydroxyphenylpyruvate, –lactate, and –acetate are excreted into the urine
(tyrosyluria).

Alkaptonuria
homogentisate 1,2-dioxygenase
is an inborn deficit of homogentisate (homogentisate oxidase)
oxidase characterized by the excretion
of homogentisate in the urine. Except for
the darkening of the urine on the air,
there are no clinical manifestations in (maleinylacetoacetate)
homogentisate
youth until the second or third decade,
when deposits of pigments in the
connective tissue begins to appear oxidation to
benzoquinoneacetate
(ochronosis – bluish colouring of the by polyphenol oxidase
scleras, the ear and nasal cartilages, or by the O2 in the air
etc.) which are the cause of deforming and polymerization to black-
arthritis. brown pigments
56
Biosynthesis of catecholamines

3-hydroxylation decarboxylation
(O2, BH4) - CO2

tyrosine DOPA
(3,4-dihydroxyphenylalanine)

β-hydroxylation N-methylation

(O2, L-ascorbate) (S-AM)

dopamine noradrenaline adrenaline

(norepinephrine) (epinephrine)

Inactivation of catecholamines occurs by means both oxidative deamination

(monoamine oxidase, MAO) to acidic metabolites and 3-O-methylation
(catechol-O-methyl transferase, COMT) to metanephrines. 57
Intermediates in the melanin biosynthesis
Pigments melanins occurs in the eye, skin, and hair. The initial steps are
a hydroxylation of tyrosine to DOPA and oxidation of DOPA to dopaquinone
– both reaction in the pigment-forming cells are catalyzed by the copper-containing
enzyme tyrosinase. The products of oxidation readily and spontaneously
undergo polymerization resulting in insoluble dark pigments.

tyrosine DOPA dopaquinone

dopachrome

indole-5,6-quinone

black eumelanins
ochre pheomelanins
58
Biosynthesis of the thyroid hormones
Within the thyroid cell, at the cell-colloid interface, iodide anions are oxidized
(to I+, IO–, or •I ?) by thyroperoxidase (TPO) and incorporated into tyrosyl
residues of thyroglobulin:

tyrosyl residues 3,5-diiodotyrosyl residues

TPO
I , H2O2
–
dehydroalanyl residue

TPO, H2O2

proteolysis

The coupling of iodotyrosyl residues iodinated thyronyl residue

in thyroglobulin is also catalyzed
by thyroperoxidase. Proteolysis of thyroxine (T4)
thyroglobulin follows in lysosomes
3,5,3´,5´-tetraiodothyronine
and thyroxine (or 3,3´,5´-T3) is secreted.
59
Tryptophan is essential and both glucogenic and ketogenic
– after opening of the indole pyrrole ring, it releases
alanine, the carbon atoms of aromatic ring give
acetoacetate.

Tryptophan mostly does not undergo transamination.

Catabolism of tryptophan is usually initiated by cleavage of the pyrrole ring
of indole by tryptophan dioxygenase (tryptophan pyrrolase):

H2O
Trp dioxygenase
O2, NADPH + H+
HCOO–
N-formylkynurenine formate kynurenine
N10-formyl TH4
pyruvate alanine

H2O hydroxylation
(O2, NADPH + H+)
(B6)

3-hydroxyanthranilate 3-hydroxykynurenin
60
 (tryptophan)

3-hydroxyanthranilate
O2

CO2
NAD+
CoA-SH CO2
hydrogenation

FAD
NH3 CO2
2-aminomuconate crotonoyl-CoA
6-semialdehyde 2-oxoadipate glutaryl-CoA
H2O hydratation
NAD
+
dehydrogenation

acetoacetyl-CoA
61
 Utilization of tryptophan

Nicotinate ring synthesis for NAD(P)+:

Humans can provide nearly all of their nicotinamide requirement from tryptophan,
if there is a sufficient amount of tryptophan in the diet. Normally, about two-thirds
comes from this source:

3-hydroxyanthranilate
O2

PRPP

H2O
PPi
CO2 ⊕
2-amino-3-carboxy- quinolinate ribosyl 5´-phosphate
muconate 6-semialdehyde
nicotinate mononucleotide
2 ATP
Gln
AMP
2 PPi
Glu

62 NAD+
 5-hydroxylation
O2 H2O

BH4 BH2
5-hydroxytryptophan CO2 serotonin
tryptophan
(5-hydroxytryptamine, 5-HT)

large intestine Serotonin is a neurotransmitter in CNS and a local hormone

decarboxylation of argentaffin cells of the intestinal mucosa. It is degraded
to 5-hydroxyindoleacetic acid (5-HIAA).
pineal gland
(N--acetylation
5-O-methylation)

tryptamine

melatonin
(N-acetyl-5-methoxytryptamine)

Secretion of melatonin from the pineal gland is increased in

indole skatole darkness. Its physiologic roles remains to be elucidated, but
they involve chronobiologic rhythms.
(In frogs, melatonin is an antagonist of the melanocyte-
stimulating hormone, MSH.)
63
The fate of the carbon skeleton of amino acids – summary:

64