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Stracture and expression of genes of the enzymes involved in purine nucleotide catabolism

Research Project

Project/Area Number 62480135
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionYokohama city university, School of medicine

Principal Investigator

TSUSHIMA Keizo  Yokohama city university, School of medicine,professor, 医学生化学, 教授 (20045917)

Co-Investigator(Kenkyū-buntansha) YAMAZAKI Kenichi  Nagoya university, School of Agriculture, research associate, 農学部生化学制御研施設 (40182480)
NISHINO Tomoko  Yokohama city university, School of medicine, research associate, 医学生化学, 助手 (80075613)
NISHINO Takeshi  Yokohama city university, School of medicine, associate professor, 医学生化学, 助教授 (40094312)
Project Period (FY) 1987 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1988: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1987: ¥4,000,000 (Direct Cost: ¥4,000,000)
KeywordsPurine nuceotide / Xanthine dehydrogenase / cDNA cloning / Structure and function of the enzyme / 反応速度論的解析 / CDNAクローニング / プリンヌクレオチド分解
Research Abstract

Previously we have purified the enzymes involved in purine catabolism to homogeneity from many sources. These enzymes are among these enzymes xanthine dehydrogenase responses most dramtaically, isolation of cDNA clone has been attempted. Furthermore, redox and kinetic properties of xanthine dehydrogenase were extensively studied.
(1) Screening of the cDNA library with rabbit antiserum to purified rat liver xanthine dehydrogenase yielded six immunologically positive clones. phage DNAs of two out of the six clones were isolated and the cDNA insert (1 kbp) of each clone was subjected to nucleotide sequence analysis. To clone longer cDNA, screening of the cDNA library were repeated.
(2) Proteolytic cleavage of the enzyme yielded several domain peptides. Most of the amino acids sequence of the smalleslt domain peptide and partial amino acid sequence were known. The NAD binding site of the enzyme was modified with 5'-FSBA. After proteolytic cleavage of the midified enzyme the modified peptide was isolated and sequenced.
(3) Kinetic studies of xanthine dehydrogenase elucidate the mechanism of the reaction of the enzyme with xanthine, NAD and oxygen as substrates.

Report

(3 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] L.M.Schopfer: The Jouranl of Biological Chemistry. 263. 13528-13538 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: The Jouranl of Biological Chemistry. 264. 3518-2527 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: The Jouranl of Biological Chemistry. 264. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: The Jouranl of Biological Chemistry. 264. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] V.Massey: The Jouranl of Biological Chemistry. 264. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y.Hattori: Advance in Experimental Medicine and Biology.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Saito: Advance in Experimental Medicine and Biology.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: Advance in Experimental Medicine and Biology.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] L.M.Schopfer: "Rapid reaction studies on the reduction and oxidation of chicken liver xanthine dehydrogenase by the xanthine/urate and NADH/NAD couples" The Journal of Biological Chemistry. 263. 13528-13538 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: "The reactivity of xanthine dehydrogenase with molecular oxygen" The Journal of Biological Chemistry. 264. 2518-2527 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: "The nicotinamide adenine dinucleotide binding site of chicken liver xanthine dehydrogenase" The Journal of Biological Chemistry. 264. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: "Reactivity of chicken liver xanthine dehydrogenase containing modified flavins" The Journal of Biological Chemistry. 264. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] V.Massey: "Difference in protein structure of xanthine dehydrogenase and xanthine oxidase revealed by reconstitution with flavin active site probes" The Journal of Biological Chemistry. 264. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Saito: "difference in redox and kinetic properties between NAD-dependent and O_2-dependent types of rat liver xanthine dehydrogenase" submitted.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y.Hattori: "Localization of xanthine dehydrogenase in liver" Advance in Experimental Medicine and Biology.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Saito: "Properties of two types of rat liver xanthine dehydrogenase" Advance in Experimental Medicine and Biology.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Nishino: "Structure of xanthine dehydrogenase" Advance in Experimental Medicine and Biology.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Saito: "Active site probe of xanthine dehydrogenase" submitted.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] L.M.Schopfer.: The Journal of Biological Chemistry. 263. 13528-13538 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Nishino.: The Journal of Biological Chemistry. 264. 2518-2527 (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Nishino.: The Journal of Biological Chemistry. 264. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Nishino.: The Journal of Biological Chemistry. 264. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] V.Massey.: The Journal of Biological Chemistry. 264. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] Y.Hattori.: Advance in Experimental Medicine and Biology.

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Saito.: Advance in Experimental Medicine and Biology. in press.

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Nishino.: Advance in Experimental Medicine and Biology.

    • Related Report
      1988 Annual Research Report
  • [Publications] K.Yamazaki;T.Nishino;K.Tsushima: Flavins and Flavoprotein(Walter de Gruy Ter,Berlin).

    • Related Report
      1987 Annual Research Report
  • [Publications] T.Saitoh;T.Nishino;K.Tsushima: Flvins and Flavoproteins(Walter de Gruyter,Berlin).

    • Related Report
      1987 Annual Research Report
  • [Publications] T.Nishion,K.Tsushima,L.Schopfer,U.Massey: Flavins and Flavoproteins(Walter de Gruyter,Boslin).

    • Related Report
      1987 Annual Research Report
  • [Publications] T.Nishion,T.Nishino;K,Tsushima;L.Schopfer;V.Massey: Flavins and Flovoprlteins(Walte de Gruy Ter,Berlin).

    • Related Report
      1987 Annual Research Report
  • [Publications] L.Schopfer,V.Massey;T.Nishino: J.Biol.Chem.

    • Related Report
      1987 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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