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Purification of L-Kynurenine 3-Monooxygenase from Pig Liver Mitochondrial Outer Membrane

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Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 398))

Abstract

L-Kynurenine 3-monooxygenase is an FAD-dependent mixed function oxidase located in the mitochondrial outer membrane (Okamoto and Hayaishi, 1969). The purification of the mammalian enzyme has been done only from rat liver mitochondria (Horn et al., 1971; Nishimoto et al., 1977; 1979) and never from other mammalian livers. In this paper we describe the method for the purification of this enzyme from pig liver mitochondrial outer membrane. It consists of (1) the extraction of the enzyme from the Chaps*-insoluble outer membrane material by the nonionic detergent, β-Octylglucoside, (2) ammonium sulfate fractionation, (3) DEAE-Sepharose CL-6B chromatography, and (4) affinity chromatography (mercurial-Sepharose).

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Reference

  • Bradford MM. 1976, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, AnalBiochem72: 248

    Article  CAS  Google Scholar 

  • Davis BJ., 1964, Disc electrophoresis. II. Method and application to human serum proteins, Ann N YAcad Sci121: 404

    Article  CAS  Google Scholar 

  • Hayaishi O., 1962, Kynurenine hydroxylase. In: Methods in Enzymology V, Colowick SP, Kaplan NO, eds., Academic Press, 807–809

    Chapter  Google Scholar 

  • Horn U, Ullrich V, Staudinger Hj., 1971, Purification and characterization of L-kynurenine 3-hydroxylase (EC 1.14.1.2.) from rat liver, Hoppe-Seyler’s Z Physiol Chem352: 837

    Article  CAS  Google Scholar 

  • Krajl M., 1965, Arapid microfluorimetric determination of monoamine oxidase, Biochem Pharmacol14: 1684.

    Article  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ., 1951, Protein measurement with Folin phenol reagent. J Biol Chem193: 265.

    CAS  Google Scholar 

  • Minamiura N, Yasunobu KT., 1978, Purification and some properties of porcine brain mitochondrial monoamine oxidase B, Biochem Pharmacol27: 2737

    Article  CAS  Google Scholar 

  • Nishimoto Y, Takeuchi F, Shibata Y., 1977, Molecular properties of L-kynurenine 3-hydroxylase from rat liver mitochondria, J Biochem1977; 81: 1413

    Google Scholar 

  • Nishimoto Y, Takeuchi F, Shibata Y., 1979, Purification of L-kynurenine 3-hydroxylase by affinity chromatography, J Chromatogr169: 357

    Article  CAS  Google Scholar 

  • Okamoto H, Hayaishi, O., 1969, Solubilization and partial purification of kynurenine hydroxylase from mitochondrial outer membrane and its electron donors. Arch Biochem Biophys131: 603

    Article  CAS  Google Scholar 

  • Parsons DF, Williams GR, Chance B., 1966, Characteristics of isolated and purified preparations of the outer and inner membranes of mitochondria, Ann N YAcad Sci1966; 137: 643

    Google Scholar 

  • Pellicciari R, Natalini B, Costantino G, Mahmoud MR, Mattoli L, Sadeghpour BM., 1994, Modulation of the kynurenine pathway in search for new neuroprotective agents. Synthesis and preliminary evaluation of (m-nitrobenzoyl) alanine, a potent inhibitor of kynurenine-3-hydroxylase. J Med Chem37: 647.

    Article  CAS  Google Scholar 

  • Uemura T, Hirai K., 1991, Kynurenine 3-monooxygenase activity of rat brain mitochondria determined by high performance liquid chromatography with electrochemical detection. In: Kynurenine and Serotonin Pathways, Schwarcz R., Young S.N. and Brown R.R., eds., Plenum Press, 531–534

    Google Scholar 

  • Youdim MBH, Collins GGS., 1971, The dissociation and reassociation of rat liver mitochondrial monoamine oxidase, Eur J Biochem18: 73

    Article  CAS  Google Scholar 

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© 1996 Plenum Press, New York

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Uemura, T., Hirai, K. (1996). Purification of L-Kynurenine 3-Monooxygenase from Pig Liver Mitochondrial Outer Membrane. In: Filippini, G.A., Costa, C.V.L., Bertazzo, A. (eds) Recent Advances in Tryptophan Research. Advances in Experimental Medicine and Biology, vol 398. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-0381-7_82

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  • DOI: https://doi.org/10.1007/978-1-4613-0381-7_82

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-8026-9

  • Online ISBN: 978-1-4613-0381-7

  • eBook Packages: Springer Book Archive

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