Amino Acid Metabolism

Hanley N. Abramson
Professor of Pharmaceutical Sciences
Wayne State University
December 2006

Incorporation of NH4 Into

Organic Compounds
-

1) NH4 + HCO3 + 2 ATP

Carbamoyl
Phosphate
Synthase I
(CPS-I)

NH2CO2PO3-2 + 2 ADP +
+
Carbamoyl Phosphate Pi + 2 H

TCA Cycle
+

2) NH4 +

Glutamate
O
O 2 CCH 2 CH2 CCO 2 - dehydrogenase

a-Ketoglutarate

NADPH +
H+

NH3+
-

O 2 CCH 2 CH2 CHCO 2 -

NADP +

Glutamate

Incorporation of NH4 Into

Organic Compounds (Cont.)
3)

NH3+
-

O 2 CCH 2 CH2 CHCO 2

Glutamate

+ NH4+ + 2 ATP
Glutamine
Synthase
Mg++

NH3+

H2 NCCH 2 CH2 CHCO 2 -

Glutamine
N of glutamine donated to other compounds
in synthesis of purines, pyrimidines,
and other amino acids

Biosynthesis of Amino Acids:

Transaminations
Amino Acid1 +a-Keto Acid2
NH3+
-

O 2 CCH 2 CH2 CHCO 2 -

Glutamate
O
O 2 CCH 2 CH2 CCO 2 -

a-Ketoglutarate

Amino Acid2 +a-Keto Acid1

O
R-CCO 2 -

Pyridoxal phosphate (PLP)Dependent Aminotransferase

NH2
R-CHCO 2 4

Transaminations: Role of PLP

CO2 H

CHO
CH2OPO3-2

HO
H3 C

CH2OPO3-2

HO

H3 C

H
-

NH3

N CHCH2CH2 CO2-

H2 O

O 2 CCH 2 CH 2 CHCO 2 -

Tautomerization

CO2-

O
-

O 2 CCH 2 CH 2 CCO 2 -

N CCH2 CH2CO2-

CH2 NH2
HO
H3 C

CH2

CH2OPO3-2

CH2 OPO3-2

HO
H2 O

H3 C

Transaminations
Glutamate
+
Pyruvate

Glutamate-Pyruvate
Aminotransferase
(Alanine Transferase ALT)

Glutamate-Oxaloacetate
Aminotransferase
(Aspartate Transferase AST)

Glutamate
+
Oxaloacetate

a-Ketoglutarate
+
Alanine
a-Ketoglutarate
+
Aspartate

Blood levels of these aminotransferases, also called transaminases,

are important indicators of liver disease
6

Metabolic Classification of
the Amino Acids
Essential

and Non-essential

Glucogenic and Ketogenic

Non-Essential Amino Acids

in Humans
Not required in diet
Can be formed from a-keto acids by
transamination and subsequent reactions
Alanine
Asparagine
Aspartate
Glutamate
Glutamine

Glycine
Proline
Serine
Cysteine (from Met*)
Tyrosine (from Phe*)

* Essential amino acids

8

Essential Amino Acids in

Humans
Required in diet
Humans incapable of forming requisite
carbon skeleton
Arginine*
Histidine*
Isoleucine
Leucine
Valine

Lysine
Methionine
Threonine
Phenylalanine
Tryptophan

* Essential in children, not in adults

9

Glucogenic Amino Acids

Metabolized to a-ketoglutarate,
pyruvate, oxaloacetate, fumarate, or
succinyl CoA
Aspartate
Asparagine
Arginine
Phenylalanine
Tyrosine
Isoleucine

Methionine
Valine
Glutamine
Glutamate
Proline
Histidine

Alanine
Serine
Cysteine
Glycine
Threonine
Tryptophan

10

Ketogenic Amino Acids

Metabolized to acetyl CoA or
acetoacetate
Isoleucine
Leucine
Threonine
Tryptophan

Lysine
Phenylalanine
Tyrosine

11

Amino Acids Formed From

a-Ketoglutarate
O
O 2 CCH 2 CH2 CCO 2 a-Ketoglutarate

4 Steps

Transamination or
Glutamate
dehydrogenase

CO2-

NH3+
-

O 2 CCH 2 CH2 CHCO 2

Glutamate
Glutamine
synthase

NH3+

H2 NCCH 2 CH2 CHCO 2 Glutamine

5 Steps

Proline
NH3+

+
H3 NCH2 CH2 CH2 CHCO 2 - Ornithine

Urea Cycle
NH2
NH3+
+
H2 N=C-HNCH2 CH2 CH2 CHCO 2 Guanidino group

Arginine
12

GABA Formation
NH3+
-

O 2 CCH 2 CH2 CHCO 2 Glutamate

Glutamate
decarboxylase

CO2

NH3+
-

O 2 CCH 2 CH2 CH2

Gamma-aminobutyrate
(GABA)

GABA is an important inhibitory neurotransmitter

in the brain
Drugs (e.g., benzodiazepines) that enhance the effects
of GABA are useful in treating epilepsy
13

Arginine Synthesis: The Urea Cycle

NH3+

N-Acetylglutamate
NHCOCH3
synthase
O 2 CCH 2 CH2 CHCO 2 O
CCH
CH
CHCO
2
2
2
2
CoASAc

Glutamate

N-Acetylglutamate
Activates

CPS-I

NH4+ + HCO3-

4 Steps

Carbamoyl
phosphate

NH2CO2PO3-2

NH3+

+
H3 NCH2 CH2 CH2 CHCO 2 Ornithine

NH3+
NH2 CONHCH2 CH2 CH2 CHCO 2 -

Ureido group

Citrulline

Ornithine
Transcarbamoylase (OTC)
(mitochondria)
14

The Urea Cycle (Contd.)

Asp
+
NH3-CHCH2CO2-

NH3+
NH2 CONHCH2 CH2 CH2 CHCO 2
Citrulline

CO2-

NH3+

+
H2 N=C-HNCH2 CH2 CH2 CHCO 2 NH-CHCH2CO2-

Arginosuccinate
synthase

CO2- Arginosuccinate

Ornithine
Transcarbamoylase
(mitochondria)

NH3+

+
H3 NCH2 CH2 CH2 CHCO 2 -

Fumarate

Urea
H2NCONH2

Arginosuccinase

Ornithine
Arginase

CO2-

NH2

-O

2C

TCA Cycle

NH3+

+
H2 N=C-HNCH2 CH2 CH2 CHCO 2 Arginine

15

Urea Formation
Occurs primarily in liver; excreted by kidney
Principal method for removing ammonia
Hyperammonemia:
Defects in urea cycle enzymes (CPS, OTC, etc.)
Severe neurological defects in neonates
Treatment:
Stop protein intake
Dialysis
Increase ammonia excretion: Na benzoate, Na
phenylbutyrate, L-arginine, L-citrulline
16

Blood Urea Nitrogen

Normal range: 7-18 mg./dL
Elevated in amino acid catabolism
Glutamate
N-acetylglutamate
CPS-1 activation
Elevated in renal insufficiency
Decreased in hepatic failure
17

Synthesis of Nitric Oxide

NH2

NH3+

Arginine
+
H2 N=C-HNCH2 CH2 CH2 CHCO 2 Nitric oxide synthase (NOS)
NH3+
NH2 CONHCH2 CH2 CH2 CHCO 2 -

NO

Citrulline

18

Nitric Oxide
Cell messenger
Implicated in a wide range of physiological
and pathophysiological events:
Vasodilation:
Activates guanylyl cyclase cGMP
Nitroglycerin
Glycerin + NO
Sildenafil (Viagra): in vascular smooth muscle:
Blocks

NO

cGMP

Phosphodiesterase-5

GMP
19

Review Questions

20

Which one of the following does

not belong on this list?
A. Carbamoyl phosphate
B. Urea
C. Glutamate
D. Glutamine

Explain your answer:

21

A glucose metabolite formed in the

transamination of alanine
___________
Derivatives of this vitamin are
involved in transamination
reactions ______________
An amino acid, not found in
peptides, that is formed in the
urea cycle ______________
The amino acid that is the
immediate precursor of urea and
NO ____________

22

Decarboxylation of ___________
affords ___________, an important

inhibitor of neurotransmission in the

CNS

Explain the importance of the Blood

Urea Nitrogen (BUN) test:
__________________________________
__________________________________
23

Categorize the following amino acids

as glucogenic (G) or ketogenic (K);
essential (E) or non-essential (NE):
Leucine: G K E NE
Phenylalanine: G K E NE
Glutamate: G K E NE

24

Formation of Serine
CO 2 -

Glucose

Dehydrogenase

Glycolysis
H C OH

NAD+ NADH +
H+
CH2 OPO3-2

3 Steps

3-Phosphoglycerate

C=O
CH2OPO3-2

3-Phosphohydroxypyruvate

Pyruvate

Inhibits
CO 2

CO 2 -

Glutamate
Transaminase
a-Ketoglutarate

CO 2 -

Phosphatase
H C NH3

CH2OH

Serine (Ser)

H C NH3 +
CH2 OPO3-2

3-Phosphoserine
25

Conversion of Serine to Glycine

H

Dihydrofolate
reductase

Folate

H2 N

CO 2 -

N
N
OH

CH 2 NHR

H C NH3+

Tetrahydrofolate
(FH4)

Serine

CH2OH

Serine hydroxymethyl
transferase (PLP-dep.)
H
N

Key intermediate
in biosynthesis of
purines and
formation of
thymine

CO 2 N

Glycine

CH 2

H C NH3
N

H2 C

N5, N10-Methylene FH4

Important in
biosynthesis of heme,
porphyrins, and purines
26

Sulfur-Containing Amino Acids

Methionine
Synthase
(Vit. B12-dep.)

NH3 +

CH 3SCH 2CH 2CHCO

- + FH4
2

NH3 +
HSCH 2CH 2CHCO 2 + 5-Methyl

FH4

L-Homocysteine

Methionine
(Essential)

CO 2 -

Cystathionine
b-synthase
(PLP-dep.)

NH3 +

OH
-

CH 3CHCH 2CO 2

b-Hydroxybutyrate

Cystathionine
lyase
-

HSCH 2CHCO 2

Cysteine
(Non-essential)

H C NH3+

Serine

CH2OH

NH3+
-

SCH 2CH 2CHCO 2

CH2CHCO2NH3+

Cystathionine
27

Homocysteine
Homocysteinuria
Rare; deficiency of cystathionine b-synthase
Dislocated optical lenses
Mental retardation
Osteoporosis
Cardiovascular disease
death
High blood levels of homocysteine associated with
cardiovascular disease
May be related to dietary folate deficiency
Folate enhances conversion of
homocysteine to methionine
28

Methionine Metabolism:
Methyl Donation
NH2
N

NH3 +

CH 3SCH 2CH 2CHCO

S-Adenosyl methionine
synthase

+
O2CCHCH2 CH2 -S-H2C

NH3 +

ATP

Methionine

+
H3 NCH2 CH2CH2-S-H2C

CO2
O2CCHCH2 CH2-S-H2C

CH3
OH OH

N
-

S-Adenosyl
Methionine
(SAM)
N

CH3

NH2

NH2

Decarboxylated
SAM

OH OH

SAM
Decarboxylase

Methyltransferases

R-CH3

NH3 +

S-Adenosyl
homocysteine

R-H

OH OH

29

Polyamine Biosynthesis
NH3+

+
H3 NCH2 CH2 CH2 CHCO 2 Ornithine
(from urea cycle)

Ornithine
decarboxylase
(ODC)
(PLP-dep.)

+
H3N
Putrescine

CO2

Decarboxylated
SAM
Spermidine
synthase

H
+
H3N

+
N

+
N
H

+
NH3

+
NH3

Spermine
synthase

Spermine
5-Methylthioadenosine

Decarboxylated
SAM

+
H3N

5-Methylthioadenosine
H
+N

+
NH3

Spermidine
30

Polyamines
Spermidine and spermine found in virtually
all procaryotic and eucaryotic cells
Precise role undefined
Bind to nucleic acids
Inhibition of biosynthetic pathway:
CO2 H
NH2
H2 N
CHF2

a-Difluoromethylornithine (DFMO)
(Eflornithine) - inhibits ODC;
used to treat
Pneumocystis carinii infectons
31

Creatine and Creatinine

NH2

Arginine-glycine
transamidinase
(Kidney)

NH3+

+
H2 N=C-HNCH2 CH2 CH2 CHCO 2 -

Glycine

Arginine

NH2

+
H2 N=C-HNCH2 CO 2 -

Ornithine

Guanidoacetate
SAM + ATP

H
N

Creatinine
(Urine)

HN

Guanidoacetate
Methyltransferase
(Liver)
Non-enzymatic
(Muscle)

N
CH3

Creatine kinase
(Muscle)

Creatine

NH2
+
H2 N=C-NCH2 CO 2 CH3

ATP

ADP
+ Pi

S-Adenosylhomocysteine
+ ADP

NHPO3-2

+
H2 N=C-NCH2 CO 2 CH3
Phosphocreatine
32

Creatine and Creatinine

Creatine:
Dietary supplement
Used to improve athletic performance
Creatinine:
Urinary excretion generally constant;
proportional to muscle mass
Creatinine Clearance Test:
Compares the level of creatinine in urine (24 hrs.)
with the creatinine level in the blood
Used to assess kidney function
Important determinant in dosing of several drugs
in patients with impaired renal function
33

Histidine Metabolism:
Histamine Formation
H
N

+
NH3
CH2 CHCO2 -

Histidine
decarboxylase

H
N

CH2 CH2 NH2

Histidine

CO2

Histamine

Histamine:
Synthesized in and released by mast cells
Mediator of allergic response: vasodilation, bronchoconstriction
(H1 receptors)
H1 blockers: Diphenhydramine (Benadryl)
Loratidine (Claritin)
Stimulates secretion of gastric acid (H2 receptors)

H2 blockers: Cimetidine (Tagamet); ranitidine (Zantac)

34

Review Questions

35

This amino acid, formed from serine using

a folate-derived coenzyme, is a key
building block in the biosynthesis of
heme and purines ________________

36

High blood levels of this amino acid, which is

not found in peptides, are associated with
increased risk of cardiovascular disease
________________
Explain why a person found to have high
levels of this amino acid might benefit from
the use of folic acid __________________

37

NH2
N

N
+
O2CCHCH2 CH2 -S-H2C

NH3 +

CH3
OH OH

What is the name of the above compound?

Explain the importance of the above
compound in the cell:

38

Name:
An arginine metabolite, formed mainly in
muscle and excreted in the urine, that is
sometimes used as a dietary supplement
to improve athletic performance ___________
An amino acid, which upon decarboxylation,
produces a compound some of whose biological
effects are blocked by cimetidine (Tagamet)
_________________
39

Phenylalanine and Tyrosine

Phenylalanine
(Essential)

H2 N

NH3+

H2O

HN

HO OH

Phenylalanine-4Monooxygenase
(Phenylalanine
hydroxylase)

NADP+
NADPH + H+

H2 N

NH3+
CH2CHCO2-

Tetrahydrobiopterin

CHCHCH3

N
H

HO

Tyrosine
(Non-essential)

H
H

CH2CHCO2O2

H
N

N
Dihydrobiopterin

+
HN
O

N
H

CHCHCH3
HO OH
40

Phenylketonuria (PKU) Disease

Deficiency of Phe hydroxylase
Occurs in 1:16,000 live births in U.S.
Seizures, mental retardation, brain
damage
Treatment: limit phenylalanine intake
Screening of all newborns mandated
in all states
Tyr
Phe
Transamination

Phenylpyruvate
(urine)

CH2CCO2-

41

Catecholamine Biosynthesis
HO
Tyr hydroxylase

NH3+

NH3+

CH2CHCO2-

O2

HO

Dihydroxyphenylalanine
(DOPA)
DOPA
decarboxylase

Epinephrine
(Adrenaline)
CHCH2NHCH3
OH

CH2CHCO2-

HO

Tyrosine
HO

Catechol

HO

Methyl
transferase

S-Adenosylhomocysteine

Dopamine
hydroxylase

HO

CH2CH2NH2

HO

HO

CO2

Dopamine

SAM

HO
DOPA, dopamine, norepinephrine,
and epinephrine are all neurotransmitters

CHCH2 NH2
OH

Norepinephrine

42

L-DOPA in Parkinsonism
Blood

L-DOPA

Brain

L-DOPA

Dopamine

HO
CH3
HO

Blocks

CH2-C-CO2H

Carbidopa

NHNH2

Dopamine

Parkinsonism associated with

dopamine in brain through loss of
neurons in basal ganglia.
Carbidopa + L-DOPA

Blood Brain Barrier

43

Monoamine Oxidase (MAO)

HO

MAO
(in mitochondria)
CHCH2NHR'

HO

HO

HO

R
OH
OH
H

R
H
Norepi
CH3 Epi
H
Dopamine

MAO inhibitors (e.g., tranylcypromine) are useful

in the treatment of depression
Brain levels of dopamine and norepi.; also
serotonin

CHCHO
R

Aldehyde
dehydrogenase
HO

HO

CHCO2 H

Urinary
metabolite

R=OH Vanillylmandelic acid (VMA)

R=H Homovanillic acid (HVA)
44

Tyramine
OH

MAO

CH2CH2NH2

OH

CH2 CHO

Tyramine
( blood pressure)
Tyramine found naturally in several types of cheese;
also beer and red wine.
Tyramine intake can cause hypertensive crisis in
persons taking a MAO inhibitor ( norepi release)
45

Catechol-O-Methyl
Transferase (COMT)
HO

HO

COMT
CHCH2NHR'
R

Active
catecholamine

HO

CHCH2NHR'

CH3O
SAM

S-Adenosylhomocysteine

COMT found in cytoplasm

Inactive
metabolite

Terminates activity of catecholamines

Catecholamine excretion products result from
combined actions of MAO and COMT
Inhibitors of COMT (e.g., tolcapone) useful
in Parkinsons disease

46

Homogentisic Acid Formation

HO

HO
NH3+

Transamination

O
CH2CCO2-

CH2CHCO2-

Tyrosine

p-Hydroxyphenylpyruvate
O2

Deficient in
alkaptonuria

Cleavage of
aromatic ring

Homogentisate
dioxygenase

CO2
O2

Fumarate + acetoacetate

OH

p-Hydroxyphenylpyruvate
dioxygenase
(ascorbate-dep.)

CH2 CO2OH

Homogentisate

47

Alkaptonuria
Deficiency of homogentisate dioxygenase
Urine turns dark on standing
Oxidation of homogentisic acid
Asymptomatic in childhood

Tendency toward arthritis in adulthood

48

Melanin Formation
HO
Tyr hydroxylase

NH3+
CH2CHCO2-

HO
NH3+

O2

Tyrosine

CH2CHCO2-

HO

DOPA
Tyrosinase
O

Melanin
(Black polymer)

Highly colored
polymeric
intermediates

Melanin formed in skin (melanocytes), eyes, and hair

In skin, protects against sunlight
Albinism: genetic deficiency of tyrosinase

CH2 CHCO2 + NH3

Dopaquinone
49

Tryptophan Metabolism:
Serotonin Formation
+
NH3

Indole ring

+
NH3

CH2 CHCO2 -

Trp
hydroxylase HO
N
H

Tryptophan
(Trp)

CH2CH2NH2

CH2 CHCO2 -

HO

Decarboxylase

O2

N
H

N
H

5-Hydroxytryptophan

CO2

5-Hydroxytryptamine (5-HT);
Serotonin

50

Serotonin
Serotonin formed in:
Brain (neurotransmitter; regulation of sleep, mood, appetite)
Platelets (platelet aggregation, vasoconstriction)
Smooth muscle (contraction)
Gastrointestinal tract (enterochromaffin cells - major storage site)

Drugs affecting serotonin actions used to treat:

Depression
Serotonin-selective reuptake inhibitors (SSRI)
Migraine
Schizophrenia
Obsessive-compulsive disorders
Chemotherapy-induced emesis

Some hallucinogens (e.g., LSD) act as serotonin agonists

51

L-Tryptophan
Food supplement promoted for serotonin effects
L-Tryptophan disaster (1989):
Eosinophilia-myalgia syndrome (EMS)
Severe muscle and joint pain
Weakness
Swelling of the arms and legs
Fever
Skin rash
Eosinophilia
Many hundreds of cases; several deaths
Traced to impurities
52

Serotonin Metabolism: 5-HIAA

CH2 CHO

CH2CH2NH2

MAO

HO

HO
N
H

N
H

Serotonin

Dehydrogenase
CH2CO2H
HO

Carcinoid tumors:
Malignant GI tumor type
Excretion of large amounts of 5-HIAA

N
H

5-Hydroxyindole acetic
acid (5-HIAA) (Urine)

53

Serotonin Metabolism:
Melatonin
CH2CH2NH2
HO

2 Steps
N
H

Serotonin

CH2 CH2 NHCOCH3

H3 CO
N
H

Melatonin

Melatonin:
Formed principally in pineal gland
Synthesis controlled by light, among other factors
Induces skin lightening
Suppresses ovarian function
Possible use in sleep disorders
54

Tryptophan Metabolism:
Biosynthesis of Nicotinic Acid
+
NH3
CH2 CHCO2 -

CO2 H

Several steps
N
H

Tryptophan

Nicotinic acid (Niacin)

Nicotinamide adenine
dinucleotide (NAD)
55

Review Questions

56

This neurotransmitter is formed by oxidation of

tryptophan, followed by decarboxylation ____________

This natural catecholamine is used as a drug to

treat Parkinsons disease ____________________.
This drugs effectiveness can be enhanced by
using ________________,
a potent decarboxylase inhibitor.

This compound is found in high levels in the blood

of patients with PKU disease ___________
57

Name the enzyme that catalyzes each of the

following reactions and explain the importance
of each reaction
HO

HO

HO

CHCH2NHR'

CH3O

CHCH2NHR'
R

HO
NH3+
HO

CH2CHCO2-

CH2 CHCO2 + NH3

58