Amino Acid Metabolism

Introduction
● AMino acids undergo oxidative degradation under three
circumstances:
○ During Normal Synthesis & breakdown of cellular protein,
when when amino acids released from normal breakdown of
cellular proteins are not needed for protein synthesis
undergo oxidative degradation.
○ When diet rich in protein are ingested and the ingested
amino acids exceeds the body’s needs for protein synthesis
the surplus is catabolised.
○ During starvation or uncontrolled diabetes mellitus when
carbohydrates are either unavailable or not properly utilised
cellular proteins are used as fuel.
● Amino acids differ from other molecules in having an amino group.
The pathways involve a key step when α-amino groups are
separated from the carbon skeleton and shunted into pathways of
amino group metabolism.
● Transamination collects amino groups of all many aminoacids in
glutamate.
● Excess ammonia generated in most other tissues is converted to
the amide nitrogen of glutamine
● In muscle amino groups are generally transferred to pyruvate to
form alanine.
Objectives
BI5.3 Describe the digestion and absorption of dietary proteins. K KH
BI5.4 Describe common disorders associated with protein metabolism. K KH
● Deamination
● Transmethylation
● Metabolism of Branched Chain Amino Acids ( BCAA)
Glutamate releases its amino group as ammonia in the liver

● In hepatocytes glutamate is transported

from cytosol from the mitochondria where
it undergoes oxidative deamination
catalysed by glutamate dehydrogenase
● It is the only enzyme that can use both
NAD+ and NADP+ as acceptor for
reducing equivalents.
● The combined effect of transamination
and deamination is known as
transdeamination.
Glutamate Dehydrogenase

● α-ketoglutarate formed from

glutamate deamination can be used
both in TCA cycle and for glucose
synthesis.
● GDH operates at an important
intersections of carbon and nitrogen
metabolism.It has ADP as a positive
modulator, and GTP as a negative
modulator.
● Mutation that alter GTP binding site
or cause otherwise permanent
activation of activity lead to a
genetic disorder known as
hyperinsulinism-
hyperammonaemia syndrome.
Oxidative Deamination by L- amino acid oxidase
● L-amino acid oxidases are FMN dependent and alongwith NH 3 also produce H2O2 , which is broken down by
catalase.
Non Oxidative Deamination
MAny amino acids undergo direct
deamination without undergoing
transamination first.
Methyltransferases

● One carbon transfers are catalysed by

Folate .
● It carries a methyl group at N5 but its
transfer potential is insufficient for most
reactions.
● S- adenosyl methionine, derived from
methionine is preferred cofactor for
biological methyl group transfers.
● The reaction is unusual in that the
nucleophilic S of methionine attacks the 5’
Carbon of of the ribose moiety of ATP rather
than one of the P atoms. Triphosphate is
released and is cleaved to Pi & PPi on the
enzyme and PPi cleaved to 2Pi by inorganic
pyrophosphatase.The methyl group is is
subject to nucleophilic attacks and is about
1000 times more reactive than CH3 group at
N5methyltetrahydrofolate
Transmethylation: Synthesis Of Creatinine & Epinephrine
Role of Vitamin B12

● In One form methionine

synthase
N5methyltetrahydrofolate is the
methyl donor.Some bacteria
and mammals utilise
N5methyltetrahydrofolate, but
the methyl group is first
transferred to cobalamin to form
methyl cobalamin as the methyl
donor.
● This reaction and the
rearrangement of L-
methylmalonyl CoA to Succinyl
CoA are the only two Vit B12
dependent reaction in man.
Role of Vitamin B12

● The conversion of Propionyl CoA a product of

odd-chain fatty acid oxidation to Succinyl CoA
requires a mutase with a derivative of cobalamin
as its coenzyme.
● Rearrangement reaction:
a. Homolytic cleavage of the C-Co bond of
5’-deoxycobalamin forming Co2+ of the
coenzyme and 5’-deoxyadenosyl radical.
b. The 5’-d-adeno radical abstracts a
hydrogen from the substrateto form a 5’-
d-adenosine and a substrate radical
c. Spontaneous rearrangement in the
substrate radical:the carbonyl CoA
migrates to the position formerly occupied
by the H forming a different radical.
d. This product radical abstracts a H from
the methyl group of the 5’-d-adenosine to
complete the rearrangement & return the
d-adenosyl unit to the radical form.
● The role of coenzyme Vit B12 is to serve as a
source of a free radical to abstract H atoms.
Alternative Fate Of Methionine
● Two alternative fates of Homocysteine:
○ Conversion to methionine (Remethylation requires Vit B12 )
○ Conversion to Cysteine(Transsulphuration, Requires Vit B6 )
● Homocysteine and vascular disease:
● Elevated levels of Homocysteine promote oxidative damage,
inflammation and endothelial dysfunction, and are independent
risk factor for occlusive vascular disease (CVD, stroke)
● Levels of plasma Hcy are inversely related to levels of folate,
B12 & B6 .Supplementation of these vitamins have been shown
to reduce levels of circulating Hcy.However in established
CVD, vitamin therapy does not reduce CVD or or even death.
● Elevated Hcy and decreased Folate in pregnant women are
associated with neural tube defects( improper closure,Spina
bifida) in fetus.Antenatal supplementation reduces the risk of
such defects.
Homocystinuria
● Large elevations of plasma Hcy as a result of rare deficiency of
cystathionine 𝛽 - synthase activity in patients with classic Homocystinuria.
● Deficiencies of the remethylation pathway also results in high levels of
homocysteine.
Branch Chain Amino
Acids (BCAA) Are Not
Degraded In The Liver
● Unlike other amino acids,BCAA are
oxidised primarily in the
muscle,adipose,kidney and brain.
● They contain a transaminase which
convert all three BCAA to
corresponding keto acid.
● Branch chain α-keto acid
dehydrogenase complex then
catalyses oxidative decarboxylation
of all three α-keto acid releasing the
carboxyl group as CO2 and forming
the acyl CoA derivative.
● Similar reactions are catalysed by
PDH complex & α-keto glutarate DH
complex. E3 is identical in all three.
Metabolic Reactions Are Plenty In No. But Limited In Types
Degradation Of Branched Chain Amino Acids
Leucine:Dehydrogenation : Isovaleryl CoA dehydrogenase

Carboxylation : similar to Acetyl CoA carboxylase

Hydration : HMG CoA → Acetyl CoA + Acetoactate

Isoleucine → Acety lCoA+ Propionyl CoA

Valine → CO2 + Propionyl CoA

Disorders: Maple Syrup urine Disease (MSUD)
● Rare autosomal recessive disorder (1: 185,000) due to partial or complete deficiency of mitochondrial enzyme complex that
oxidatively decarboxylates BCAA.
● The three α-keto acids, as well as their precursor amino acid, leucine accumulates in the blood and spillover in the urine and
gives the characteristic odour in the urine imparted by the α-keto acids.
● Diagnosis can be made by treating urine with 2,4 dinitrophenyl hydrazine which gives 2,4 dinitrophenyl hydrazone. Definitive
diagnosis by MS.
● Feeding problems, vomiting, ketoacidosis, change in muscle tone,neurological problem leading to coma( primarily due to
increase in leucine).
● Untreated disease is fatal. If treatment is delayed, intellectual disability results.
● Variants :
○ Classic : Neonatal onset,most common form, absent or little BCKD activity.Letha within the first few weeks if untreated.
○ Intermediate : Higher level of enzyme activity ( upto 30%), symptom molder & onset in infancy to adolescence.
○ Thiamine dependent :Respond to large doses of vitamin.
● Treatment : With a synthetic formula free of BCAA ,supplemented with limited amount of Leu, Ile, Val. Rarely diagnosis and life
long treatment essential.
● Note: BCAA are an important energy source, and MSUD patient are at risk of decompensation during periods of increased
protein catabolism.
Isovaleric Acidemia
● Cherry like smell of breath and body fluids( sweat, Urine)on ingestion of
excessive protein diet.
● The odour is due to accumulation of Isovaleryl CoA due to Isovaleryl CoA
deficiency.
Methylmalonic Aciduria
● Methyl malonyl CoA is the intermediate in the catabolism of valine. Its normal
fate is its conversion to succinyl CoA by isomerase requiring Vit B12 as
coenzyme.
● Hence, in some cases associated with Vit B12 deficiency, marked improvement
is seen with Vit B12.
● A more severe form of seen that requires much higher doses of Vit B12 upto
1G/day.
Inherited defects of Amino Acid Metabolism
Metabolic
Disorders