Amino Acid Metabolism MS1031

Dr Andrew K Snabaitis

Amino Acid Metabolism

Source of metabolic energy during starvation
Precursors:
-muscle protein metabolised into keto acids (gluconeogenesis)

- structural proteins - hormones - lipids - ketone bodies

Energy production via oxidative metabolism

- late stage glycolysis - TCA cycle

Amino Acid Metabolism

Metabolic Classes
20 aas are needed for protein synthesis - non essential aas endogenously synthesized - essential aas (10) cannot be endogenously synthesized

Amino Acid Metabolism

Metabolic Classes
Subclassification - glucogenic: aas converted to glucose - ketogenic: oxidised to acetyl-CoA or acetoacetate and then ketone bodies

Amino Acid Metabolism

Absorption
Dietary protein is hydrolysed to: - free aas - peptides (2 or 3 aas) - transported into intestinal epithelial cells (enterocytes) - hydrolysed to free aas - transport into bloodstream (portal vein) Enterocytes - glutamine (Gln) preferred energy source - glutamate (Glu) and aspartate (Asp) secondary fuel source - alanine (Ala) produced forms glucose via gluconeogenesis Alanine - liver extracts alanine from bloodstream and removes amino nitrogen by transamination to reform pyruvate

Amino Acid Metabolism

Metabolism of nitrogen in amino acids
- nitrogen component (a-amino group) removed before complete metabolism - transamination: transfer to suitable a-keto acceptor - deamination: oxidative removal to form oxo acids and release ammonia (NH3)

Transamination (liver)

- reversible reaction catalysed by aminotransferases - removed amino groups can be used to synthesize new non-essential aas - aminotransferases are specific for particular aas - glutamate from 2-oxoglutarate - aspartate from oxaloacetate - pyruvate can accept amino gp to form alanine - malate can accept amino gp to form glutamine - hence Ala/Glu transport amino gps from tissues to liver

Amino Acid Metabolism

Transamination (skeletal muscle)
- post-absorptive state alanine and glutamine are released from skeletal muscle - >50% of total aas released - other aas metabolised to give pyruvate and 2-oxoglutarate - form alanine from pyruvate - form glutamine from 2-oxoglutarate - alanine and glutamine released into bloodstream via aa membrane transporters

Amino Acid Metabolism

Transamination (skeletal muscle)
- branched chain aas (leucine, isoleucine and valine) major source of nitrogen for alanine and glutamine production - branched chain a-keto aas released into bloodstream are taken up by liver and synthesizes: - glucose from keto acid of valine - ketone bodies from keto acid of leucine - glucose and ketone bodies from keto acid of isoleucine

Amino Acid Metabolism

Transamination (Clinical implications)
Transaminase enzymes - plasma transaminase enzymes increase due to liver damage - aspartate aminotransferase (AST) is a mitochondrial enzyme - alanine transaminase (ALT) is a cytosolic enzyme - used as clinical markers of liver damage/disease - AST found in heart, skeletal muscle, kidney and brain
Vitamin B6 deficiency - transamination requires pyridoxal phosphate (cofactor) derived from vitamin B6 - drugs (isoniazid) interact with pyridoxal phosphate - peripheral neuropathy, anaemia and dermatitis - vitamin B6 supplements reverse conditions

Amino Acid Metabolism

Deamination
- oxidative removal of a-amino gp from aa by oxidases - keto acids and NH3 released - glutamate is oxidatively deaminated by glutamate DH to form: - 2-oxoglutarate - ammonia (NH3) enters the urea cycle - NH3 in blood diffuses across BBB - NH3 uptake by brain causes glutamate to form glutamine glutamate deficiency - glutamate used to form g-aminobutyric acid (GABA) - GABA is an important inhibitory neurotransmitter - linked to encephalopathy

Deamination (clinical implications)

Amino Acid Metabolism

Urea cycle
- urea synthesis takes place in liver - both mitochondrial and cytosolic reactions - carbamoyl phosphate is synthesized from NH3, CO2 and ATP in hepatic mitochondria by carbamoyl phosphate synthetase (CPS-1) - ornithinec transported to mitochondria to form citrulline (condensation) - citrulline diffuses into cytosol to form argininosuccinate (condensation)

Amino Acid Metabolism

Urea cycle (contd)
fumerate
- argininosuccinate split into arginine and

- fumerate used to regenerate aspartate - arginine split by arginase to form urea - aspartate condensed with citrulline - urea transported in blood to be excreted by kidneys in urine - urea cycle intersects with TCA cycle - some fumerate forms malate via fumerase and then oxaloacetate via malate DH - oxaloacetate transaminated to aspartate by aspartate aminotransferase

Amino Acid Metabolism

Urea cycle (regulation)
- short term: allosteric CPS-1 activator (N-acetylglutamate) - long term: gene expression of enzymes of urea cycle

Glucagon and glucocorticoids regulate CPS-1 gene expression CPS-1 gene expression increases: - high-protein diets - starvation - proteins used for energy

Amino Acid Metabolism

Urea cycle defects (Clinical implications)
CPS-1 deficiency - severe UCD - hyperammonaemia causes vomiting, poor feeding, convulsions, ataxia and coma - fatal if not rapidly diagnosed post partum Other enzyme deficiencies - ornithine transcarbamoylase - argininosuccinate synthetase - argininosuccinase - arginase Treatment of UCD - restrict dietary protein - removal of NH3 from GI tract - antibiotics e.g. benzoate/phenylacetate

Amino Acid Metabolism

Essential amino acids - valine, leucine and isoleucine represent 15-20% dietary protein

- 70% of total aas entering blood stream after a meal - 60-80% of total aas taken up by muscle - surplus not used for protein synthesis used to generate energy

Essential amino acids (clinical implications) - phenylketonuria (PKU) - 1 in 20,000 births in UK and USA - tyrosine synthesis is abrogated due to phenylalanine hydroxylase deficiency - excrete phenylpyruvate and phenylacetate in urine - develop brain damage, mental retardation and epilepsy - tyrosine supplements are successful - alkaptonuria (black urine disease) - deficiency of homogentisic acid oxidase - very rare 1 in 1,000,000 births - phenylalanine and tyrosine metabolism generates homogentisic acid metabolite - excreted in urine that turns black/brown on standing - pigment can accumulate in joints and cause arthiritis

Amino Acid Metabolism

Essential amino acids (clinical implications) - maple syrup urine disease - rare condition (1 in 300,000 births) - defect in keto acid decarboxylase - accumulation of keto acids derived from branched chain aas - give urine a maple syrup odour - infant epilepsy, neonatal acidosis, brain damage and death - no treatment - albinism - tyrosinase deficiency - tyrosinase hydroxylates tyrosine to dihydroxyphenylalanine (DOPA) - DOPA is a precursor of skin pigment melanin - amelanosis: lack of skin, hair and iris colouration - no neurological disturbances - benign condition

Amino Acid Metabolism

Amino acids as signalling molecules - glutamate and glycine act directly as neurotransmitters - glutamate: excitatory neurotransmitter in brain - glycine: inhibitory neurotransmitter in spinal cord - other aas undergo metabolism to form neurotransmitters - tyrosine: adrenaline, noradrenaline and dopamine - tryptophan: serotonin - arginine & leucine implicated in activation of mTOR

Core Texts
Medical Sciences (1st Ed) Naish, Revest & Syndercombe Court
Medical Biochemistry (3rd Ed) Baynes & Dominiczak