METABOLISM OF PROTEINS

AND
AMINO ACIDS
Revised by Levi Letlet H. Larcia II, MS
Adapted from Ph Ch 127 slides prepared by Joanna J. Orejola, MS
OUTLINE

• Overview
• Digestion and Absorption of Proteins and Transport
of Amino Acids
• Amino Acid Deamination
• Urea Cycle and Metabolic Fate of Nitrogen
• Catabolism of Amino Acids
• Amino Acid Biosynthesis
• Amino Acids as Biosynthetic Precursors
OVERVIEW

• Role of Amino
Acids in the Body
• Amino Acid Pool
• Protein Turnover
• Consequence of
Protein Imbalance
Role of Amino Acids in the Body

•Precursors for nitrogen-containing

compounds
•Building blocks for protein synthesis
•Source of energy
Amino Acid Pool • RDA 56g/day for a 70
kg man
• primary source of the
• 100 g amino acids nitrogen metabolized
• Sources: by the body
• Dietary protein
• Body protein
• Synthesis of nonessential amino acids

• ~400g/day
• Only 75% of hydrolysis
product are recaptured
through the biosynthesis
of new tissue protein
 Protein Turnover
• constantly synthesized and degraded
• In healthy humans:
• rate of protein synthesis is just sufficient to
replace degraded protein
• Leads to hydrolysis and resynthesis of
300-400g body protein/ day
• Rate of turnover
• varies widely for individual proteins
• Proteins have different half-lives
•PEST proteins
• have short half-lives; hypothesized to act as a
signal peptide for protein degradation
Consequence of Protein Imbalance
• Too much protein

Excess
amino acid

Glucose ammonia
or fat
Consequence of Protein Imbalance
• Too little protein:
Kwashiorkor Marasmus
DIGESTION AND ABSORPTION OF
PROTEINS AND TRANSPORT OF
AMINO ACIDS
DIGESTION AND ABSORPTION OF PROTEINS
AND TRANSPORT OF AMINO ACIDS

• Protein Digestion
• Protein Absorption
• Amino Acid Transport
Protein Digestion
• starts in the stomach,
completed in the intestine
• Enzymes:
• produced as inactive precursors
• secreted in the digestive tract
lumen
• proteolysis
• have different specificities:
• no single enzyme can digest dietary
protein
 Protein Digestion
Digestion by Pancreatic Enzymes

procarboxy-
trypsinogen chymotrypsinogen proelastase peptidase A & B
endopeptidase
carboxy-
trypsin chymotrypsin elastase peptidase A & B

R O R O R O R O R O
+ NH NH NH NH -
H3N NH NH NH NH O
NH NH
O R O R O R O R
O R
Arg Trp Ala Ala
Lys Tyr Gly Ile
Phe Ser A Leu
Met Val
Leu Arg
B
Lys
Protein Digestion

Digestive Enzymes Digest:

• dietary proteins themselves
• intestinal cells regularly sloughed off into the lumen
Protein Absorption

• Intestinal epithelia:
• Free amino acids
• entering the portal system readily
• dipeptides
• hydrolyzed first in the cytosol before entering the
portal system
Amino Acid Transport
Mechanisms
• Secondary Active Na+-dependent Transport
• Facilitated Diffusion
• Transport-Linked to the Glutamyl Cycle
Amino Acid Transport
Secondary Active Na+-dependent
Intestinal
at least 6 different Lumen
types of
Brush Border
transporters
Active
apical brush Transporter
border membrane
ATP
have overlapping Pi +
specificity for ADP
Facilitated
different amino transporter
acid Serosal side
also present in the
Na+
renal epithelium Amino acid
Portal vein
K+
 Amino Acid Transport
Facilitated Transport Intestinal
•out of the cells Lumen

through the serosal Brush Border

side Active
•bidirectional Transporter

ATP
Pi +
ADP
Facilitated
transporter
Serosal side

Na+
Portal vein
Amino acid
K+
Amino Acid Transport
ADP + Pi ATP
Linked to the
Glutamyl Cycle γ-glutamyl-
• across the intestine cysteinylglycine
glycine
γ-glutamylcysteine

and kidney ADP + Pi

cysteinylglycine
membranes
cysteine ATP

γ-glutamylamino acid

glutamate
5-oxoproline

ATP ADP + Pi

Amino acid

γ-glutamyl-transpeptidase
Amino Acid Transport

•Cystinuria
• genetic error in the transporter protein:
• Cystine, Lysine, Arginine, and Ornithine
• will not suffer from amino acid deficiency
• precipitation of cystine forming kidney stones

•Hartnup Disease
• defect in transport of neutral amino acids across both
intestinal and renal epithelial cells
• first observed in the Hartnup Family
• amino acid deficiency is expected
Amino acid deamination
AMINO ACID DEAMINATION

• Transamination
• Oxidative Deamination
Transamination
- -
O O
+ O
NH3
O O
R
R
amino acid α-keto acid
aminotransferases

PLP
- - - -
O O O O

O O O O
+
O NH3

α-ketoglutarate glutamate
 Oxidative Deamination
• Glutamate: O H O

• Mitochondrion -
O
-
O + NAD(P)+

• glutamate NH3
+

dehydrogenase- glutamate

• the only enzyme that O O

can accept either NAD+ -
O O
-
+ NAD(P)+H + H+
or NADP+ as coenzyme
• Inhibited in vitro by
+
NH2
α-iminoglutarate
GTP H2O

• activated by GDP & O O

ADP O
- -
O
+ NH4
+

O
α-ketoglutarate
UREA CYCLE AND METABOLIC
FATE OF NITROGEN
UREA CYCLE AND METABOLIC
FATE OF NITROGEN

• Urea Cycle
• Metabolic Fate of Nitrogen
 Mitochondrion

2ATP O
2ADP + Pi
H2N C OPO32-
NH3 + HCO3- Carbamoyl phosphate
Carbamoyl phosphate
synthetase
Pi
O Ornithine O
O
transcarbamoylase
+ -
H3N O C -
+ H2N NH O
NH3 +
Citrulline NH3
Ornithine

O
Urea -
O -
O O

H2N C NH2 Urea Cycle ATP O NH3

Aspartate
+

Arginase
AMP + PPi Argininosuccinate
O synthetase
H2O Cytosol -
O -
O
H2N
O O HN
O
+
C - C
H2N NH O -
+ NH O
+ fumarate H2N
+
Arginine NH3
Argininosuccinate NH3
Argininosuccinase
Urea Cycle

Regulation
•Allosteric activation by N-acetylglutamate
• Activates CPS I
•Substrate availability
• “feed forward” mechanism
• With the exception of arginase, deficiency in urea
cycle enzymes results to substrate accumulation
•Induction/repression of enzyme synthesis
• Dependent on rate of protein metabolism
Metabolic Fate of Nitrogen
Sources of NH4+ for Urea Cycle Aspartate
Purine
Nucleotide
Cycle Fumarate
glutamate α-ketoglutarate MUSCLE
GDH

NAD+ NADH PLP

Serine Pyruvate
Threonine
glutamine glutamate NH4+ PLP
α-ketobutyrate

asparagine aspartate
Histidine Urocanate
Urea bacteria
Amino acids
Various
GUT products
Metabolic Fate of Nitrogen

Transport of Ammonia in NH2 O

the Circulation O OH

• urea GLUTAMINE
NH2

• from the liver to the H2O ADP + Pi

kidneys glutamine synthetase
• passes into the glomerular NH4
+
ATP + NH4
+

filtrate
• Glutamine O O

• provides a nontoxic storage HO OH

and transport form of NH2
ammonia GLUTAMATE
 Metabolic Fate
of Nitrogen

Modes of Nitrogen Excretion

• Ureotelic
• Ammonotelic
• uricotelic
CATABOLISM OF AMINO ACIDS
 CATABOLISM OF AMINO ACIDS
• Glucogenic and Ketogenic Amino Acids
• Degradation of Ala, Cys, Gly, Ser and Thr to Pyruvate
• Degradation of Asn and Asp to Oxaloacetate
• Degradation of Arg, Glu, Gln, His, and Pro to α-
ketoglutarate
• Degradation of Ile, Met and Val to Succinyl-CoA
• Degradation of Leu, Ile, and Val to Acetoacetate
and/or Acetyl-CoA
• Degradation of Lys to Acetoacetate and/or Acetyl-
CoA
• Degradation of Trp to Alanine and Acetoacetate
• Degradation of Phe and Tyr to Fumarate and
Acetoacetate
CATABOLISM OF AMINO
ACIDS
Glucogenic and
Ketogenic Amino Acids
• Amino acids are degraded to
their citric cycle
intermediates or their
precursors so that they can
be metabolized to CO2 and
H2O or used in
gluconeogenensis
CATABOLISM OF
AMINO ACIDS

Glucogenic
• carbon skeletons are degraded to
pyruvate , α-ketoglutarate, succinyl-
CoA, fumarate or oxaloacetate
• glucose precursors
CATABOLISM OF AMINO
ACIDS

Ketogenic
• carbon skeletons are broken
down to acetyl CoA or
acetoacetate
• can be converted to fatty acids
or ketone bodies
Degradation of
Ala, Cys, Gly Ser
and Thr to
Pyruvate

1-alanine aminotransferase
2-serine dehydratase
3-glycine cleavage system
4&5-serine hydroxymethyl transferase
6-threonine dehydrogenase
7-α-amino-β-ketobutyrate lyase
H2S, SO32-, SCN-
H3C—C—COO-

O
Pyruvate
Degradation of Asn and Asp to Oxaloacetate
Degradation of
Arg, Glu, Gln, His,
and Pro to -
ketoglutarate

1-glutamate dehydrogenase
2-glutaminase
3-arginase
4-ornithine-δ-
aminotransferase
5-glutamate semialdehyde
dehydrogenase
6-proline oxidase
7-spontaneous
8-histidine ammonia lyase
9-urocanate hydratase
10-imidazolone propionase
11-glutamate formimino
transferase
 H

Degradation of Ile,
Met and Val to
Succinyl-CoA
1-methionine adenosyl transferase
2-methylase
3-adenosylhomocysteinase
4-homocysteine methyltransferase
5-cystathionine β-synthase
6-cystathionine lyase
7-α-keto dehydrogenase
8-propionyl –CoA carboxylase
9-methylmalonyl-CoA racemase
10-methylmalonyl-CoA mutase
Degradation of Leu,Ile and
Val to Acetoacetate and/or
Acetyl-CoA
1-branched chain amino acid amino
transferase Maple
2-branched chain α-keto acid Syrup
dehydrogenase
Disease
3-acyl-CoA dehydrogenase
4-enoyl-CoA hydratase
5-β-hydroxyacyl-CoA dehydrogenase
6-acetyl-CoA acetyl transferase
7-enoyl-CoA hydratase
8- β-hydroxyisobutyryl-CoA hydrolase
9- β-hydroxyisobutyrate dehydrogenase
10-methylmalonate semialdehyde
dehydrogenase
11-β-methylcrotonyl-CoA-carboxylase
12- β-methylglutaconyl-CoA hydratase
13-HMG-CoA lyase
 Degradation of Lys to
Acetoacetate and/or
Acetyl-CoA

1-saccharopine dehydrogenase
2-saccharopine dehydrogenase
3-aminoadipate semialdehyde
dehydrogenase
4-aminoadipate aminotransferase
5-α-keto acid dehydrogenase
6-glutaryl-CoA dehydrogenase
7-decarboxylase
8-enoyl-CoA hydratase
9-β-hydroxyacyl-CoA dehydrogenase
10-HMG-CoA synthase
11-HMG-CoA lyase
Degradation
of Trp to Ala and
Acetoacetate
1-tryptophan-2-3-
dioxygenase
2-formamidase
3-kynurenine-3-
monooxygenase
4-kynureninase
5-3-hydroxyanthranilate-
3,4-dioxygenase
6-aminocarboxy
muconate
7-aminomuconate
semialdehyde
dehydrogenase
8-hydratase
9-dehydrogenase
Degradation of Phe and
Tyr to Fumarate and
Acetoacetate

1-phenylalanine hydroxylase
2-aminotransferase
3-p-hydroxyphenylpyruvate
dioxygenase
4-homogentisate dioxygenase
5-maleylacetoacetate isomerase
6-fumarylacetoacetase
AMINO ACID BIOSYNTHESIS
AMINO ACID BIOSYNTHESIS

• Pyruvate as Precursor
• Oxaloacetate as Precursor
• α-Ketoglutarate as Precursor
• 3-Phosphoglycerate as Precursor
• Glutamate as Precursor
Pyruvate as Precursor
Oxaloacetate as Precursor
α-Ketoglutarate as
Precursor
 3-Phosphoglycerate
as Precursor
1-3-phosphoglycerate dehydrogenase
2-aminotransferase
3-phosphoserine phosphatase
Glutamate as
Precursor
1-γ-glutamylkinase
2-dehydrogenase
3-nonenzymatic
4-pyrroline-5-carboxylate
reductase
5-N-acetylglutamate
synthase
6-acetylglutamate kinase
7-N-acetyl-γ-glutamyl
phosphate dehydrogenase
8-N-acetylornithine-δ-
aminotransferase
9-acetylornithie deacetylase
10-ornithine-δ-
aminotransferase
AMINO ACIDS AS BIOSYNTHETIC
PRECURSORS
• Biosynthesis of Neurotransmitters and other N-
containing Compounds
Biosynthesis of Neurotransmitters
and other N-containing Compounds
END OF LECTURE