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1987 Fiscal Year Final Research Report Summary

Study on ganglioside syntheses

Research Project

Project/Area Number 61440089
Research Category

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

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionHokkaido University

Principal Investigator

MAKITA Akira  Hokkaido University School of Medicine, 医学部, 教授 (60004561)

Co-Investigator(Kenkyū-buntansha) KASI Noriyuki  Hokkaido University School of Medicine, 医学部, 助教授 (60001947)
HONKE Kouichi  Hokkaido University School of Medicine, 医学部, 助手 (80190263)
GASA Shinsei  Hokkaido University School of Medicine, 医学部, 助教授 (10142712)
Project Period (FY) 1986 – 1987
KeywordsGanglioside GM1 synthase / GM3 synthase / GD3 synthase / Galactosyltransferase / Sialyltransferase / シアリルトランスフェラーゼ / スルフォトランスフェラーゼ
Research Abstract

1. GM_2 synthase which catalyzes the conversion of ganglioside GM_3 to GM_2 was purified over 6300-fold from rat liver particulate fractions by varius chromatographies. The purified enzyme consisted of two identical subunits of 64,000 dalton. GM_2 containing either NeuAc or NeuGc and the oligosaccharide, sialyllactose were good substrates for GM_3 synthase, indicating that the preferred acceptor has a structure ofNeuAc(or NeuGc) 2-3GalBl-4Glc-OR.
2. GM_1 synthase which forms GM_1 from GM_2 was purified from rat liver Golgi apparatus by affinity chromatographies to a 10,2000-fold. The synthase acted partly (28% of GM_2) on Gg_3Cer, but not on other glycolipids and glycoproteins tested.
3. We have developed a new method for assay of GM_3 aynthase, which forms GM_3 from LacCer, using a DEAE-Sephadex column. Using this method, GM_3 synthase was partialy purified from particulate fraction of pig spleen by conventinal and affinity chromatographies.
4. GD_3 synthase which forms GD_3 from GM_3 was fractionated from rat liver Golgi apparatus by conventional and affinity chromatographies. Factors and conditions affecting the synthase were determined.
5. Cerebroside sulfotransferase was purified from rat kidney, in parallel with the above studies. The purified enzyme had a wide range of substrate specificity acted on LacCer (110%) and Gg_3Cer (85%) in addition to Gal cer (100%), indicating that the same enzyme sulfates these glycolipids.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Honke Kouichi: Anal. Biochem.155. 395-399 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yanagisawa Ken: Biochim. Biophys. Acta. 919. 213-220 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishikawa Yukitoshi: J. Biochem.101. 1369-1375 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sako Fumiyo: Int. J. Biochem.19. 923-929 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Gasa Shinsei: J.Biol. Chem.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano Masatugu: Anal. Biochem.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Honke Kouichi: "A radioimmune-assay of ganglioside GM_1 synthase using cholera toxin" Anal. Biochem.155. 395-399 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yanagisawa Ken: "Purification and properties of GM2 synthase, UDP-N-acetylgalactosamine: GM3 -N-acetylgalactosaminyltransferase from rat liver" Biochim. Biophys. Acta. 919. 213-220 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishikawa Yukitoshi: "Characterization of neutral glycosphingoloipids from fetal human brain. Evidence for stage-specific expression of the globo, ganglio and neolacto series in the central nervous system" J. Biochem.101. 1369-1375 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sako Fumiyo: "Characterization of neutral glycosphingolipids from porcine erythrocyte membranes" Int. J. Biochem.19. 923-929 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Gasa Shinsei: "Purification and characterization of cerebroside sulfotransferase from rat kidney" J. Biol. Chem.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawano Masatugu: "A rapid assay method for glycosphingolipid synthases using ion-exchange chromatography" Anal. Biochem.

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

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Published: 1989-03-30  

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