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Identification of L-amino acid/L-lysine α-amino oxidase in mouse brain

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Abstract

Lysine, an essential amino acid is catabolized in brain through only the pipecolic acid pathway. During the formation of pipecolic acid, α-deamination of lysine, and the formation of the α-keto acid as well as its cyclized product are pre-requisites. The enzyme mediated α-deamination of L-lysine and the formation of the α-keto acid and the cyclized product are not demonstrated so far. Both lysine and pipecolic acid are known to increase in brain under the conditions of fasting, studies were therefore undertaken to identify the enzyme responsible for the α-deamination of L-lysine in the brain tissue of mice which were fasted. The detection of the α-keto acid of L-lysine, α-keto-ε-amino caproic acid and its cyclized product, Δ 1-piperidine-2-carboxylate was facilitated by the use of L-[U-14-C]-lysine as the substrate. The quantitation of the radioactivity in reaction products was done after separation by ion exchange chromatographic methods. The formation of the α-keto acid was enzyme mediated, the α-keto acid formed was established by reaction with N-methyl benzothiazolinone hydrazone hydrochloride. The cyclized product was accounted in a fraction which matched the resolution of authentic pipecolic acid on the Dowex column, and the cyclized product was confirmed by spectrophotometry. The hitherto undemonstrated α-amino deaminating enzyme of L-lysine in brain tissue, the α-keto acid of L-lysine and its cyclized product in a mammalian system could thus be demonstrated in the present study. These findings confirm the involvement of L-lysine oxidase/L-amino acid oxidase in the formation of pipecolic acid from L-lysine.

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References

  1. Meister A: Intermediary Metabolism of Amino Acids. In: Biochemistry of Amino Acids, 2nd edn. Academic Press, New York, Vol. 11 1965, pp 928–1020

    Google Scholar 

  2. Scriver CR, Rosenberg LE: Lysine. In: Amino Acid Metabolism and its Disorders. Saunders, Philadelphia, 1973, pp 250–255

    Google Scholar 

  3. Higashino K, Tsukada K, Lieberman J: Saccharopine, a product of lysine breakdown in mammalian liver. Biochem Biophys Res Commun 20: 285–290, 1965

    Google Scholar 

  4. Higashino K, Fujioka M, Akoi T, Yamamura Y: Metabolism of lysine in rat liver. Biochem Biophys Res Commun 29: 95–100, 1967

    Google Scholar 

  5. Higashino K, Fujioka M, Yamamura Y: The conversion of L-Lysine to saccheropine and α-amino adipate in mouse. Arch Biochem Biophys 142: 606–614, 1971

    Google Scholar 

  6. Hutzier J, Dancis J: Conversion of lysine to saccharopine by human tissues. Biochem Biophys Acta 158: 62–69, 1968

    Google Scholar 

  7. Chang YF: Lysine metabolism in the rat brain: The pipecolic acid forming pathway. J Neurochem 30: 347–354, 1978

    Google Scholar 

  8. Dagley S, Nicholson DE: Amino Acids in An Introduction to Metabolic Pathways. Blackwell Science Publishing, Oxford, 1970, pp 190–249

    Google Scholar 

  9. Chu SW, Hegsted DM: Adaptive response of lysine and threonine degrading enzymes in adult rats. J Nutrition 106: 1089–1096, 1976

    Google Scholar 

  10. Torchinsky Yu M: Transamination: Its discovery, biological and chemical aspects. (1937–1987) Trends Biochem Sci 12: 115–117, 1987

    Google Scholar 

  11. Broquist P: Lysine-Pipecolic Acid relationship in microbes and mammals. In: Ann. Review of Nutrition 11 1991 pp 435–448

    Google Scholar 

  12. Rodwell VW: Catabolism of Carbon Skeletons of Amino Acids. In: R.K. Murray, D.K. Graner, P.A. Mayes, V.W. Rodwell (eds). Harpers Biochemistry, 23rd edn. Prentice Hall Int. Inc., 1993, pp 303–325

  13. Boulanger E, Bertrand J, Oesteux R: Desamination de l'orinithine et de LA lysine selectivement marquees par la L-amino acide-dehydrogenase du foie de dindou (Meleagris Calloparo L). Biochem Biophys Acta 26: 143–145, 1957

    Google Scholar 

  14. Nishio H, Sawda E, Sogabe H, Segawa T: Effect of starvation and immobilization on amino acids in mouse brain and peripheral tissues. Neurochem Int 8(2): 229–233, 1986

    Google Scholar 

  15. Weismann N, Schoenheimer R: The relative stability of L(+)-Lysine in rats studied with deuitirium and heavy nitrogen. J Biol Chem 140: 779–795, 1941

    Google Scholar 

  16. Rothstein M, Miller LL: The conaversion of L-lysine-6–14C to pipecolic acid in the rat. J Am Chem Soc 75: 4371–4372, 1953

    Google Scholar 

  17. Rothstein M, Miller LL: The conversion of L-lysine to pipecolic acid in the rat. J Biol Chem 211: 851–858, 1954

    Google Scholar 

  18. Schmidt-Glenewinkel T, Nomura Y, Glacobini E: The conversion of lysine into piperidine, cadaverine, and pipecolic acid in the brain and other organs of the mouse. Neurochem Res 2: 619–637, 1977

    Google Scholar 

  19. Giacobini E, Nomura Y, Schmidt-Glenewinkel T: Pipecolic acid: Origin, biosynthesis and metabolism in the brain. Cell Mol Biol 26: 135–145, 1980

    Google Scholar 

  20. Gatfield PD, Taller E, Hinton GG, Wallace AC, Abdelnoum GM, Haust MD: Hyperpipecolatemia: A new metabolic disorder associated with neuropathy and hepatomegaly. Can Med Assoc J 99: 1215–1233, 1968

    Google Scholar 

  21. Thomas GH, Hasiam RH, Batshaw ML, Caputa AJ, Neidengard L, Raonsom JL: Hyperpipecolic acidemia associated with hepatomegaly, mental retardation, optic nerve dysplasia and progressive neurological disease. Clin Genet 8: 376–382, 1975

    Google Scholar 

  22. Trijbels JMF, Monnens LAH, Bakkeren JAJAM, Raay Selten AHJ, Corsteaensen JMB: Biochemical studies in the cerebro-hepato-renal syndrome of Zellweger: A disturbance in the metabolism of pipecolic acid. J Inher Metab Dis 2: 39–42, 1979

    Google Scholar 

  23. Burton BK, Reed SP, Remy WT: Hyperpipecolic acidemia: Clinical and biochemical obsersvations in two male siblings. J Pediat 99: 729–734, 1981

    Google Scholar 

  24. Chalia VR, Geisinger KR, Burton BK: Pathological alteration in the brain and liver in hyperpipecolic acidemia. J Neuropath Exp Neurol 42: 627–638, 1983

    Google Scholar 

  25. Kelley RI, Moser HW: Hyperpipecolic aciduria in neonatal adrenoleukodystrophy. Am J Med Genet 19: 791–795, 1984

    Google Scholar 

  26. Kase Y, Takahama K, Hashimoto T, Kaisaku J, Okano Y, Miyata T: Electrophoretic study of pipecolic acid a biogenic imino acid, in the mammalian brain. Brain Res 193: 608–613, 1980

    Google Scholar 

  27. Nomura Y, Okuma Y, Segawa T, Schmidt-Glenewinkwel T, Giacobini E: Comparison of synaptosomal and glial uptake of pipecolic acid and GABA in rat brain. Neurochem Res 6: 391–400, 1981

    Google Scholar 

  28. Giacobini E: Imino acids of the brain. In: A. Lajtha (ed). Hand book of Neurochemistry, 2nd edn. Raven Press, Vol. 3, 1983, pp 583–605

  29. Feigenbaum P, Chang YF: Pipecolic acid antagonizes barbiturate enhanced GABA binding to bovine brain membranes. Brain Res 372: 176–179, 1986

    Google Scholar 

  30. Meister A, Radhakrishnan AN, Buckley SD: Enzymatic synthesis of L-pipecolic acid and L-proline. J Biol Chem 229: 789–800, 1957

    Google Scholar 

  31. Murthy SN, Janardana Sarma MK: Enzymatic conversion of L-lysine to α-keto ∈-amino caproic acid. (Abstr) J Neurochem 63 (Suppl.1): S28 D, 1994

    Google Scholar 

  32. Kusakabe H, Kodama K, Kuninaka A, Yashino H, Misono H, Soda K: A new antitumor enzyme, L-lysine α-oxidase from Trichoderma viride. J Biol Chem 255: 976–981, 1980

    Google Scholar 

  33. Lajtha A: In: A Lajtha (ed). Handbook of Neurochemistry. 2nd edn. Vol IV. 1983, pp 77–110

  34. Murthy SN, Sarma MKJ: Detection of the cyclised product of α-keto acid of L-lysine-precursor for the formation of pipecolic acid. (Abstract), Annual Meeting of Society of Biological Chemists (India) 1996

  35. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254, 1976

    Google Scholar 

  36. Soda K: Microdetermination of D-amino acids and D-amino acid oxidase activity with 3–methyl-2–benzothiazolone hydrazone hydrochloride. Anal Biochem 25: 228–235, 1968

    Google Scholar 

  37. Rodwell VW: Pipecolic acid. In: H. Tabor, C.W. Tabor (eds). Methods in Enzymology XVII B, 1971, pp 174–188

  38. Katsoyannis PG, Schwartz GP: The synthesis of peptides by homogenous solution procedures. In: C.H.W. Hirs, S.W. Timasheff (eds). Methods in Enzymology XLVII, 1979, pp 501–578

  39. Knott RJ, Joseph MH, Curzon G: Effects of food deprivation and immobilisation on tryptophan and other amino acids in rat brain. J Neurochem 20: 249–280, 1973

    Google Scholar 

  40. Volpe JJ, Lee G, Laster L, Robinson JC: Regional distribution of isoenzymes of D-amino acid oxidase and acetyl esterases in developing primate brain. Exp Neurol 28: 76–87, 1970

    Google Scholar 

  41. Weliner D, Lichtenberg LA: Assay of amino acid oxidase. In: H. Tabor, C.W. Tabor (eds). Methods in Enzymology XVII B, 1971, pp 593–596

  42. Ratner S: A long view of nitrogen metabolism. In: E.E. Snell, P.D. Boyer, A. Meister, C.L. Richardson (eds). Ann Rev Biochem 46: 1977, pp 1–24

  43. Soda K, Misono H: L-lysine α-ketoglutarate amino transferase (Achromobacter liquidum) In: H. Tabor, C.W. Tabor (eds). Methods in Enzymology XVII B, 1971, pp 222–223

  44. Rao DR, Rodwell VW: Metabolism of pipecolic acid in a Pseudomonas species. J Biol Chem 237: 2232–2238, 1962

    Google Scholar 

  45. Basso LV, Rao DR, Rodwell VW: Metabolism of pipecolic acid in a Pseudomonas species. J Biol Chem 237: 2239–2245, 1962

    Google Scholar 

  46. Paz MA, Blumendfeld OO, Rojkind M, Henson E, Furfine C, Gallop PM: Determinantion of carbonyl compounds with N-methyl benzothiazolone hydrazone. Arch Biochem Biophys 109: 548–559, 1965

    Google Scholar 

  47. Hamilton PB: Ion exchange chromotography of amino acids. Anal Chem 35: 2055–2064, 1963

    Google Scholar 

  48. Lehninger A: Oxidative degradation of amino acids. In: Biochemistry. Worth Publushing Inc., New York, 1970, pp 433–454

    Google Scholar 

  49. Wickwire BM, Wagner C, Broquist HP: Pipecolic acid biosynthesis in Rhizoctonia leguminicola. J Biol Chem 265: 14748–14753, 1990

    Google Scholar 

  50. Fellows FCI, Carson NAJ: Enzyme studies in a patient with saccharopinuria: A defect in lysine metabolism. Pediat Res 8: 42–49, 1974

    Google Scholar 

  51. Meister A: General Biochemical and Physiological considerations. In: Biochemistry of Amino Acids, 2nd edn. Academic Press, New York. Vol 1, 1965a, pp 269–437

    Google Scholar 

  52. Hagihara H, Hayashi H, Ichihara A, Suda M: Metabolism of L-lysine by bacterial enzymes. J Biochem (Tokyo), 48: 267–276, 1960

    Google Scholar 

  53. Bixell G, Hamprecht B: Generation of ketone bodies from leucine by cultured astroglial cells. J Neurochem 65: 2450–2461, 1995

    Google Scholar 

  54. Pevzner: Multiple forms of enzymes. In: A. Lajtha (ed). Handbook of Neurochemistry, Vol. IV, 2nd edn. 1983, pp 461–484

  55. Hill JM: Diamine oxidase (pea seedling). In: H. Tabor, C.W. Tabor (eds). Methods in Enzymology XVII B, 1971, pp 730–735

  56. Neuberger A, Sanger F: The availability of the acetyl derivatives of lysine for growth. Biochem J 37: 515–518, 1943

    Google Scholar 

  57. Neuberger A, Sanger F: The metabolism of lysine. Biochem J 58: 119–125, 1944

    Google Scholar 

  58. Meister A: Enzymatic preparation of α-Keto acids. J Biol Chem 197: 309–317, 1952

    Google Scholar 

  59. Hernandez MF, Chang YF: In vitro synthesis of L-pipecolate from Llysine: Inconsistent with ∈-acetyl L-lysine as an obligatory intermediate. Biochem Biophys Res Commun 93: 762–769, 1980

    Google Scholar 

  60. Murthy SN: Ph.D. Thesis - Studies on the enzymatic conversion of Llysine to pipecolic acid: Identification of L-amino acid oxidase in mouse brain. Osmaania University, Hyderabad, India, 1996

    Google Scholar 

  61. Greenstsein JP, Birnbaum SM, Otey MC: Optical and enzymatic characterization of amino acids. J Biol Chem 204: 307–321, 1953

    Google Scholar 

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  1. Department of Neurochemistry, National Institute of Nutrition, Jamai Osmania, Hyderabad, India

    S.N. Murthy & M.K. Janardanasarma

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  1. S.N. Murthy
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Murthy, S., Janardanasarma, M. Identification of L-amino acid/L-lysine α-amino oxidase in mouse brain. Mol Cell Biochem 197, 13–23 (1999). https://doi.org/10.1023/A:1006906505745

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