TRANSAMINATION & DEAMINATION

TRANSAMINATION & DEAMINATION

1. Amino acid Metabolism Gandham.Rajeev Email:[email protected]

2. Proteins are the most abundant organic compounds & constitute a major part of the body dry weight (10-12kg in adults). Perform a wide variety of structural & dynamic (enzymes, hormones, clotting factors, receptors) functions. Proteins are nitrogen containing macromolecules consisting of L- α - amino acids as the repeating units.

3. Of the 20 amino acids found in proteins, half can be synthesized by the body & half are supplied through diet. The proteins on degradation release individual amino acids. Each amino acid undergoes its own metabolism & performs specific functions.

4. Some amino acids serve as precursors for the synthesis of many biologically important compounds. Certain amino acids may directly act as neurotransmitters ( e.g glycine, aspartate, glutamate)

5. Amino acid pool About 100g of free amino acids which represent the amino acid pool of the body. Glutamate & glutamine together constitute about 50% & essential amino acids about 10% of the body pool ( 100g). The concentration of intracellular amino acids is always higher than the extracellular amino acids. Enter the cells against a concentration gradient.

6. Sources of amino acid pool Turnover of body intake of dietary protein & the synthesis of non-essential amino acids contribute to the body amino acid pool. Protein turnover: The proteins in the body is in a dynamic state. About 300-400g of protein per day is constantly degraded & synthesized which represents body protein turnover.

7. Control of protein turnover: The turnover of the protein is influenced by many factors. A small polypeptide called ubiquitin (m.w.8,500) tags with the protein & facilitates degradation. Certain proteins with amino acid sequence proline , glutamine, serine & threonine are rapidly degraded.

8. Dietary protein: There is a regular loss of nitrogen from the body due to degradation of amino acids. About 30-50g of protein is lost every day. This amount of protein is supplied through diet to maintain nitrogen balance. There is no storage form of amino acids in the body. Excess intake of amino acids is oxidized to provide energy.

9. U tilization of amino acids from body pool Proteins function as enzymes, hormones, immunoproteins , contractile proteins etc. Many important nitrogenous compounds ( porphyrins , purines , pyrimidines , etc) are produced from the amino acids. About 10-15% of body energy requirements are met from the amino acids. The amino acids are converted into carbohydrates & fats.

10. Catabolism of amino group occurs in 4 stages Transamination Oxidative Deamination Ammonia Transport Urea Cycle

11. Transamination The transfer of an amino (-NH2) group from an amino acid to a ketoacid , with the formation of a new amino acid & a new keto acid. Catalysed by a group of enzymes called transaminases ( aminotransferases ) Pyridoxalphosphate (PLP)– Co-factor. Liver, Kidney, Heart, Brain - adequate amount of these enzymes.

12. Salient features of transamination All transaminases require PLP. No free NH3 liberated , only the transfer of amino group. Transamination is reversible. There are multiple transaminase enzymes which vary in substrate specificity. AST & ALT make a significant contribution for transamination.

13. Transamination is important for redistribution of amino groups & production of non-essential amino acids. It diverts excess amino acids towards the energy generation. Amino acids undergo transamination to finally concentrate nitrogen in glutamate.

14. Glutamate undergoes oxidative deamination to liberate free NH3 for urea synthesis . All amino acids except, lysine, threonine, proline & hydroxyproline participate in transamination. It involves both anabolism & catabolism, since – reversible.

15. AA 1 + α - KG ketoacid 1 + Glutamate Alanine + α - KG Pyruvate + Glutamate Aspartate + α - KG Oxaloacetetae + Glutamate

16. Glutamate-collecting point

17. Transamination

18. Step:1 Transfer of amino group from AA 1 to the coenzyme PLP to form pyridoxamine phosphate. Amino acid1 is converted to Keto acid2 . Step:2 Amino group of pyridoxamine phosphate is then transferred to a keto acid 1 to produce a new AA 2 & enzyme with PLP is regenerated. Mechanism of transamination

19. Clinical Significance Enzymes, present within cell, released in cellular damage into blood. ↑ AST - Myocardial Infarction (MI). ↑ AST, ALT – Hepatitis, alcoholic cirrhosis. Muscular Dystrophy.

20. The amino group of most of the amino acids is released by a coupled reaction , trans-deamination. Transamination followed by oxidative deamination . Transamination takes place in the cytoplasm. Trans-deamination

21. The amino group is transported to liver as glutamic acid, which is finally oxidatively deaminated in the mitochondria of hepatocytes.

22. The removal of amino group from the amino acids as NH3 is deamination. Deamination results in the liberation of ammonia for urea synthesis. The carbon skeleton of amino acids is converted to keto acids. Deamination may be either oxidative or non-oxidative Deamination

23. Only liver mitochondria contain glutamate dehydrogenase ( GDH) which deaminates glutamate to α - ketoglutarate & ammonia. It needs NAD + as co-enzyme. It is an allosteric enzyme. It is activated by ADP & inhibited by GTP.

24. Oxidative Deamination Oxidative deamination is the liberation of free ammonia from the amino group of amino acids coupled with oxidation. Site: Mostly in liver & kidney. Oxidative deamination is to provide NH3 for urea synthesis & α - keto acids for a variety of reactions , including energy generation.

25. Role of glutamate dehydrogenase Glutamate is a 'collection centre ' for amino groups. Glutamate rapidly undergoes oxidative deamination. Catalysed by GDH to liberate ammonia . It can utilize either NAD + or NADP + . This conversion occurs through the formation of an α- iminoglutarate

26. Oxidation of glutamate by glutamate by GDH COO - I CH2 I CH2 I H - C- NH 3 + I COO - COO - I CH2 I CH2 I C= NH I COO - COO - I CH2 I CH2 I CH2 I C= O I COO - NAD(P)+ GDH H 2 O GDH + NH 4 + L-Glutamate α - Iminoglutarate α - ketoglutarate NAD(P)H+H +

27. Metabolic Significance Reversible Reaction Both Anabolic & Catabolic. Regulation of GDH activity: Zinc containing mitochondrial, allosteric enzyme. Consists of 6 identical subunits. Molecular weight is 56,000.

28. Allosteric regulation GTP & ATP – allosteric inhibitors. GDP & ADP - allosteric activators. ↓ Energy - ↑ oxidation of A.A. Steroid & thyroid hormones inhibit GDH.

29. L-amino acid oxidase & D-Amino acid oxidase. Flavoproteins & Cofactors are FMN & FAD . Act on corresponding amino acids to produce α - keto acids & NH3 Site: Liver, kidney, Peroxisomes. Activity of L-Amino acid o xidase is low. Plays a minor role in Amino acid catabolism. Amino acid Oxidases

30. L-amino acid α - keto acid + NH 3 L-amino acid oxidase FMN FMNH 2 H 2 O 2 ½ O 2 Catalase H 2 O Oxidative deamination of amino acids

31. L-Amino acid Oxidase acts on all Amino acids, except glycine & dicarboxylic acids. Activity of D-Amino oxidase is high than that of L-Amino acid oxidase D-Amino oxidase degrades D-Amino acids in bacterial cell wall.

32. Fate of D-amino acids D-amino acids are found in plants & microorganisms. They are not present in mammalian proteins. D-amino acids are taken in the diet/bacterial cell wall, absorbed from gut - D-Amino acid oxidase converts them to respective α - keto acids.

33. The α - ketoacids undergo transamination to be converted to L-amino acids which participate in various metabolic pathways . Keto acids may be oxidized to generate energy or serve as precursors for glucose & fat synthesis.

34. Metabolic fate of D-amino acids D-amino acid α - Ketoacid L-amino acid Glucose & Fat FAD FADH 2 D-amino acid oxidase H 2 O NH 4 + L-Amino acid α - Ketoacid Transaminases Energy

35. Direct deamination, without oxidation. Amino acid Dehydratases : Serine, threonine & homoserine are the hydroxy amino acids. They undergo non-oxidative deamination catalyzed by PLP-dependent dehydratases Non-Oxidative deamination

36. Serine Threonine Homoserine Respective Ketoacid Dehydratase NH 3 PLP Non-Oxidative deamination

37. Cysteine & homocysteine undergo deamination coupled with desulfhydration to give keto acids. Deamination of histidine : Cysteine Pyruvate NH 3 +H 2 S Desulfhydrases Histidine Uroconate Histidase NH 3 A mino acid Desulfhydrases

38. References Textbook of Biochemistry-U Satyanarayana Textbook of Biochemistry-DM Vasudevan

39. Thank You