Purification and Properties of Pyridoxal Kinase from Bovine Brain

  • Tae Hirakawa-Sakurai
  • Kiyoshi Ohkawa
  • Makoto Matsuda
Part of the Developments in Molecular and Cellular Biochemistry book series (DMCB, volume 9)


A 27, 000-fold purification of pyridoxal kinase from bovine brain tissue has been achieved by a combination of ammonium sulfate fractionation, DEAE-cellulose chromatography, hydroxyapatite chromatography, Sephadex G-150 gel filtration, Blue Sepharose CL-6B chromatography, and Phenyl-Superose chromatography. The final chromatography step yields a homogeneous preparation of high specific activity (2105 nmol/min/mg protein). The molecular mass of the native enzyme was estimated to be approximately 80, 000 on gel filtration. The subunit 39, 500. This indicates that pyridoxal kinase is a dimeric enzyme. (Mol Cell Biochem 119: 203–207, 1993)

Key words

pyridoxal kinase bovine brain vitamin B6 


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  1. 1.
    McCormick DB, Gregory ME, Snell EE: Pyridoxal phosphoki-nases. I. Assay, distribution, purification, and properties. J Biol Chem 236: 2076–2084, 1961PubMedGoogle Scholar
  2. 2.
    McCormick DB, Snell EE: Pyridoxal phosphokinases II. Effects of inhibitors. J Biol Chem 236: 2085–2088, 1961PubMedGoogle Scholar
  3. 3.
    White RS, Dempsey WB: Purification and properties of vitamin B6 kinase from Escherichia coli B. Biochemistry 9: 4057–4064, 1970PubMedCrossRefGoogle Scholar
  4. 4.
    Neary JT, Diven WD: Purification, properties, and a possible mechanism for pyridoxal kinase from bovine brain. J Biol Chem 245: 5585–5593, 1970PubMedGoogle Scholar
  5. 5.
    Kwok F, Churchich JE: Brain pyridoxal kinase: Purification, substrate specificities, and sensitized photodestruction of an essential histidine. J Biol Chem 254: 6489–6495, 1979PubMedGoogle Scholar
  6. 6.
    Cash CD, Maitre M, Rumigny JF, Mandel P: Rapid purification by affinity chromatography of rat brain pyridoxal kinase and pyridoxamine-5-phosphate oxidase. Biochem Biophys Res Commun 96: 1755–1760, 1980PubMedCrossRefGoogle Scholar
  7. 7.
    Kerry JA, Rohde M, Kwok F: Brain pyridoxal kinase: Purification and characterization. Eur J Biochem 158: 581–585, 1986PubMedCrossRefGoogle Scholar
  8. 8.
    Tagaya M, Yamano K, Fukui T: Kinetic studies of the pyridoxal kinase from pig liver: Slow-binding inhibition by adenosine tetra-phosphopyridoxal. Biochemistry 28: 4670–4675, 1989PubMedCrossRefGoogle Scholar
  9. 9.
    Sakurai T, Asakura T, Matsuda M: Transport and metabolism of pyridoxine and pyridoxal in mice. J Nutr Sci Vitamimol 33:11–19, 198Google Scholar
  10. 10.
    Sakurai T, Asakura T, Matsuda M: Transport and metabolism of pyridoxine in the intestine of the mouse. J Nutr Sci Vitaminol 34: 179–187, 1988PubMedCrossRefGoogle Scholar
  11. 11.
    Sakurai T, Asakura T, Mizuno A, Matsuda M: Absorption and metabolism of pyridoxamine in mice I. Pyridoxal as the only form of transport in blood. J Nutr Sci Vitaminol 37: 341–348, 1991PubMedCrossRefGoogle Scholar
  12. 12.
    Sakurai T, Asakura T, Mizuno A, Matsuda M: Absorption and metabolism of pyridoxamine in mice II. Transformation of pyridoxamine to pyridoxal in intestinal tissues. J Nutr Sci Vitaminol 38: 227–233, 1992PubMedCrossRefGoogle Scholar
  13. 13.
    Wada H, Snell EE: The enzymatic oxidation of pyridoxine and pyridoxamine phosphates. J Biol Chem 236: 2089–2095, 1961PubMedGoogle Scholar
  14. 14.
    Kurioka S, Ishioka N, Sato J, Nakamura J, Ohkubo T, Matsuda M: Assay of vitamin Bb6 in human plasma with graphitic carbon column. Anal Biochem, submittedGoogle Scholar
  15. 15.
    Weber K, Osborn M: The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem 244: 4406–4412, 1969.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 1993

Authors and Affiliations

  • Tae Hirakawa-Sakurai
    • 1
  • Kiyoshi Ohkawa
    • 1
  • Makoto Matsuda
    • 1
  1. 1.Dept. BiochemJikei UnivMinato-KuTokyoJapan

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