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1998 Fiscal Year Final Research Report Summary

Enzyme system responsible for protein lipoylation

Research Project

Project/Area Number 09670132
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionThe University of Tokushima

Principal Investigator

FUJIWARA Kazuko  the University of Tokushima, the Institute for Enzyme Research, assistant professor, 分子酵素学研究センター, 助教授 (20108880)

Project Period (FY) 1997 – 1998
Keywordslipoyltransferase / lipoate-activating enzyme / lipoic acid / chromosomal mapping / glycine cleavage system / alpha-ketoacid dehydrogenase complex / cloning
Research Abstract

Lipoic acid is a prosthetic group of H-protein of the glycine cleavage system and the acytransferase components of the pyruvate, alpha-ketoglutarate, and branched chain alpha-ketoacid dehydrogenase complexes. In mammals, lipoic acid is activated to lipoyl-AMP by lipoate-activating enzyme and then the lipoyl moiety is transferred to the proteins by the lipoyltransferase.
1) We cloned a lipoyltransferase cDNA from a bovine liver cDNA library and determined the nucleotide sequence. The predicted amino acid sequence showed 35% identity with that of Escherichia coli lipoate-protein ligase A.It was suggested that lipoyltransferase I and II, isoforms found in bovine liver, were derived from the same translated products but processed differently.
2) Lipoyltransferase had an affinity for selenolipoyl-AMP and transfered the selenolipoyl moiety to bovine apoH-protein comparable to lipoyl-AMP.Reaction rates of overall and partial glycine cleavage reactions with selenolipoylated and lipoylated H-protein were compared. The results well reflected the difference of redox potential between diselenide bond and disulfide bond.
3) We cloned human lipoyltransferase cDNA and genomic DNA.Tissue distribution of the mRNA of lipoyltransferase showed good corelation with those of lipoate-dependent proteins, suggesting a possibility of an efficient lipoylation of the proteins in the tissues.
4) We have partially purified lipoate-activating enzyme from bovine liver mitochondria. The research for the enzyme is now in progress.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Kazuko Fujiwara: "Synthesis and characterization of selenolipoylated H-protein of the glycine cleavage system" J.Biol.Chem.272. 19880-19883 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuko Fujiwara: "Cloning and expression of a cDNA encoding bovine lipoyltransferase" J.Biol.Chem.272. 31974-31978 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuko Fujiwara: "Molecular cloning and expression of cDNA encoding lipoyltransferase from bovine liver" The FASEB Journal. 11. A883 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤原 和子: "ウシとヒトのリポ酸転移酵素のcDNAクローニングと発現" 生化学. 69. 581 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 藤原 和子: "ヒトのリポ酸転移酵素の遺伝子クローニングと構造解析" 生化学. 70. 913 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuko Fujiwara: "Molecular cloning,structural characterization and chromosomal localization of human lipoyltransferase gene" Eur.J.Biochem.(in press). (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuko Fujiwara: "Synthesis and characterization of selenolipoylated H-protein of the glycine cleavage system" J.Biol.Chem.272. 19880-19883 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuko Fujiwara: "Cloning and expression of a cDNA encoding bovine lipoyltransferase" J.Biol.Chem.272. 31974-31978 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuko Fujiwara: "Molecular cloning and expression of cDNA encoding lipoyltransferase from bovine liver" The FASEB Journal. 11. A883 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuko Fujiwara: "Cloning and expression of cDNAs encoding bovine and human lipoyltransferases" Seikagaku. 69. 581 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuko Fujiwara: "Molecular cloning and structural characterization of human lipoyltransferase gene" Seikagaku. 70. 913 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuko Fujiwara: "Molecular cloning, structural characterization and chromosomal localization of human lipoyltransferase gene" Eur.J.Biochem.(in press).

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-12-08  

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