Other amino acids form
succinyl CoA
• Branched-chain amino acids,
isoleucine, leucine and valine are
essential amino acids
• Metabolized primarily by the
peripheral tissues
• Undergo transamination,
oxidative decarboxylation and
dehydrogenation
Amino acids that form acetyl
CoA or aceto-acetyl CoA
• Leucine, isoleucine, lysine, and tryptophan
form acetyl CoA or aceto-acetyl CoA directly,
without pyruvate serving as an intermediate
• Phenylalanine and tyrosine also give rise to
acetoacetate
Overview of N₂ metabolism
• Biosynthetic pathways of amino acids and
nucleotides require nitrogen
• Few species can convert atmospheric nitrogen
into forms useful to living organisms
• Certain prokaryotes can fix atmospheric nitrogen
• The first important product of nitrogen fixation is
ammonia utilizable by all organisms
• Carried out by a highly conserved complex of
proteins called the Nitrogenase complex
Nitrogenase
complex
• Consists of 2 components
dinitrogenase reductase and
dinitrogenase
• Nitrogen fixation is carried out
by a highly reduced form of
dinitrogenase
• It is reduced by the transfer of
electrons from dinitrogenase
reductase
• ATP hydrolysis overcomes the
high activation energy of
nitrogen fixation
Assimilation of NH₃ into amino acids
• Amino acids, glutamate and glutamine provide the
critical entry point.
• Via transamination and action of glutamate
dehydrogenase
• Glutamate is the source of amino groups for most
other amino acids, through transamination reactions
• Amide nitrogen of glutamine is a source of amino
groups in a wide range of biosynthetic processes
• Important pathway catalysed by glutamine
synthetase
Glutamine synthetase regulation
• Primary regulatory point in nitrogen metabolism
• Central metabolic role as an entry point for reduced
nitrogen
• Regulated both allosterically and by covalent modification
• The effects of multiple allosteric inhibitors are more than
additive
• Superimposed on the allosteric regulation is inhibition by
adenylylation (addition of AMP)
• Adenylylation and deadenylylation promoted by
adenylyltransferase
Allosteric regulation of
glutamine synthetase
Second level of regulation of glutamine
synthetase:
covalent modifications. (a) An
adenylylated Tyr residue. (b) Cascade
leading to adenylylation (inactivation)
PII-UMP mediates the
activation of transcriptio
of the gene encoding
glutamine synthetase
Covalent modification of glutamine
synthetase
• This covalent modification increases the enzyme's sensitivity to the allosteric inhibitors,
and the enzyme's activity decreases as more of the 12 subunits are adenylylated.
• Both adenylylation and deadenylylation are promoted by the enzyme adenylyl
transferase, part of a complex enzymatic cascade that responds to levels of glutamine,
α-ketoglutarate, ATP, and Pi.
• The activity of adenylyl transferase is modulated by binding to a regulatory protein
called PII.
• The effect of PII, in turn, is regulated by covalent modification (uridylylation), again at a
Tyr residue.
• The adenylyl transferase complex with PII-UMP stimulates deadenylylation, whereas the
same complex with deuridylylated PIIstimulates adenylylation of glutamine synthetase.
• The uridylylation and deuridylylation of PII is brought about by a single enzyme, uridylyl
transferase, with both uridylylation and deuridylylation activities. Uridylylation is
stimulated by α-ketoglutarate and ATP but inhibited by glutamine and Pi. The
deuridylylation activity is not regulated.
• The net result of this complex mechanism is a decrease in glutamine synthetase activity
when glutamine levels are high and an increase in activity when glutamine levels are
low and the α-ketoglutarate and ATP substrates are available.
Classes of reactions occurring in synthesis of
amino acids
1. Transamination reactions
2. Transfer of one-carbon groups
3. Transfer of amino groups derived from the
amide nitrogen of glutamine
• Glutamine amido-transferases catalyse transfer
of amide groups from glutamine
Biosynthesis of
Amino acids(NE)
• Derived from intermediates
in glycolysis, TCA cycle, or
PPP
• Nitrogen enters these
pathways by way of
glutamate and glutamine
• Mammals can synthesize
only half of 20 AA
• Nonessential amino acids vs
essential amino acids
α-Ketoglutarate gives rise to Glutamate,
Glutamine, Proline & Arginine
• Proline is a cyclized derivative of glutamate
• Arginine is synthesized from glutamate via
ornithine and the urea cycle in animals
Biosynthesis of proline
glutamate kinase + glutamate
dehydrogenase
Non-enzymatic
pyrroline carboxylate
reductase
Serine, Glycine, and Cysteine are derived
from 3-Phosphoglycerate
• 1st step is an oxidation of
-OH of 3-PG
• Transamination from
glutamate yields 3
-phosphoserine, which is
hydrolyzed to free serine
• Removal of a carbon atom
yields glycine
• Can also be formed from a
reaction catalysed by glycine
synthase
Mammals synthesize cysteine
from 2 amino acids: methionine
provides sulfur atom & serine
furnishes the C-skeleton
Biosynthesis of cysteine from
homocysteine and serine in
mammals. The homocysteine is
formed from methionine
• Alanine and aspartate are synthesized from
pyruvate and oxaloacetate by transamination
from glutamate
• Asparagine is synthesized by amidation of
aspartate
• Tyrosine can be synthesized in 1 step from
phenylalanine
Regulation of AA bio-
synthesis
• Allosterism
feedback inhibition of the first reaction in a sequence
by the end product of the pathway
concerted inhibition/cumulative feedback inhibition
enzyme multiplicity
Metabolic defects in Amino
acid metabolism
• Commonly caused by mutant genes that
result in abnormal proteins, most often
enzymes
• Results in mental
retardation/developmental
abnormalities
• Most of these are generally rare
Phenylketonuria
• Caused by a deficiency of:
phenylalanine hydroxylase
enzymes that synthesize or
reduce the coenzyme
tetrahydrobiopterin
• BH₄ is also required for action
of tyrosine hydroxylase &
tryptophan hydroxylase
• Hyperphenylalaninemia
• Hypopigmentation
• CNS symptoms
Albinism
• Defect in tyrosine
metabolism results in a
deficiency in the
production of melanin
• Absence of pigment from
the skin, hair & eyes
• Results from a deficiency
of tyrosinase activity
Albinism in animals
