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Glycosylation of Membrane Sterol Associated with Cell Differentiation

Research Project

Project/Area Number 63580114
Research Category

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

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

Principal Investigator

MUROFUSHI Kimiko  Ochanomizu University, Department of Biology, Assistant, 理学部, 助手 (00103557)

Project Period (FY) 1988 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1989: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1988: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsPhysarum polycephalum / Cell differentiation / Poriferasterol monoglucoside / UDP-glucose:poriferasteol glucosyltransferase / Sterol / Phospholipids / Phospholipase D / ポリフェラステロ-ルモノグルコシド / UDP-グルコ-ス:ポリフェラステロ-ルグルコシルトランスフェラ-ゼ / ホスフォリパ-ゼD / 真性粘菌 / ポリフェラステロールモノグルコシド / UDP-グルコース / ボリフェラステロールグルコシルトランスフェラーゼ / 分化
Research Abstract

Haploid myxoamoebae of Physarum polycephalum conjugate in pairs to form diploid zygotes and differentiate into plasmodia. During the course of differentiation, an expression of a nonpolar glycoconjugate in membrane fraction was detected. This substance was purified and structural studies were performed by GC, GC/MS, FAB/MS, NMR and enzymatic hydrolysis, ant the substance has been identified as poriferasterol monoglucoside (published in J. Biol. Chem. 262, 16719-16723).
UDP-glucose:poriferasterol glucosyltransferase which synthesizes poriferasterol monoglucoside was also expressed during a differentiation from myxoamoebae to plasmodia. The enzyme activity was not detected in the amoeboid cells, but after conjugation, the enzyme activity appeared and increased. The enzyme was partially purified and characterized, and its molecular weight was determined to be 72,000, and pH optimum was 7.0. It catalyzed the glucose transfer from UDP-glucose to sitosterol and campesterol as well as to poriferasterol, but stigmasterol which is C-24 epimer of poriferasterol was a poor substrate for this enzyme reaction. Apparent Km values were 1.2 x 10^<-4>M for UDP-glucose and 4.8 x 10^<-6>M for poriferasterol, respectively. (Biochim. Biophys. Acta 992, 412-415).
Colonia strain of Physarum polycephalum is a mutant which differentiates from myxoamoebae to plasmodia without nuclear DNA change. The poriferasterol monoglucoside and its synthesizing enzyme were detected in both stages, myxoamoebae and plasmodia, in this mutant strain.
Changes in phospholipid composition and phospholipase D activity were also observed during a differentiation from myxoamoebae to diploid plasmodia in both, wild-typed and mutant strains (Biochim. Biophys. Acta,in press). Sterol contents were examined and compared, then the major one was determined to be poriferasterol (Cell Struct. Funct. 12, 519-524).

Report

(3 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Minowa,A.: "Changes in phospholipid composition and phospholipase D activity during the differentiation of a true slime mold,Physarum polycephalum." Biochim.Biophys.Acta. inpress (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Murakami-Murofushi,K.: "Expression of UDP-glucose:poriferasterol glucosyltransferase in the process of differentiation of a true slime mold,Physarum polycephalum." Biochim.Biophys.Acta. 992. 412-415 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Murakami-Murofushi,K.: "Sterol content of haploid myxoamoebae of Physarum polycephalum.Comparison with theose of diploid plasmodia." Cell Struct.Funct.12. 519-524 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Murakami-Murofushi,K.: "Expression of poriferasterol monoglucoside associated with differentiation of PHysarum polycephalum." J.Biol.Chem.262. 16719-16723 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Minowa, A., Kobayashi, T., Shimada, Y., Maeda, H., Murakami-Murofushi, K., Ohta, J., and Inoue, K.: "Changes in phospholipid composition and phospholipase D activity during the differentiation of Physarum polycephalum." Biochim. Biophys. Acta, in press (1990).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Murakami-Murofushi, K., and Ohta, J.: "Expression of UDP-glucose:poriferasterol glucosyltransferase in the process of differentiation of a true slime mold, Physarum polycephalum." Biochim. Biophys. Acta 992, 412-415 (1989).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Murakami-Murofushi, K., Nakamura, K., Ohta, J., and Yokota, T.: "Sterol content of haploid myxoamoebae of Physarum polycephalum. Comparison with those of diploid plasmodia." Cell Struct. Funct., 12, 519-524 (1987).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Murakami-Murofushi, K., Nakamura, K., Ohta, J., Suzuki, M., Suzuki, A., Murofushi, H., and Yokota, T.: "Expression of poriferasterol monoglucoside associated with differentiation of Physarum polycephalum." J. Biol. Chem., 262, 16719-16723 (1987).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Minowa,A.: "Changes in phospholipid composition and phospholipase D activity during the differentiation of Physarum polycephalum." Biochim.Biophys.Acta. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Murakami-Murofushi,K.: "Expression of UDP-glucose:poriferasterol glucosyltransferase in the process of defferentiation of a true slime mold,Physarum polycephalum." Biochim.Biophys.Acta. 992. 412-415 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Murakami-Murofushi,K.: "Sterol content of haploid myxoamoebae of Physarum polycephalum.Comparison with those of diploid plasmodia." Cell.Struct.Funct.12. 519-524 (1987)

    • Related Report
      1989 Annual Research Report
  • [Publications] Murakami-Murofushi,K.: "Expression of poriferasterol monoglucoside associated with differentiation of Physarum polycephalum." J.Biol.Chem.262. 16719-16723 (1987)

    • Related Report
      1989 Annual Research Report
  • [Publications] Kimiko ;Murakami-Murofushi: Biochim.Biophys.Acta. (1989)

    • Related Report
      1988 Annual Research Report

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Published: 1988-04-01   Modified: 2016-04-21  

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