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GTP-binding Proteins in Starfish Oocyte

Research Project

Project/Area Number 03044063
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionTokyo Instiute of Technology

Principal Investigator

KATADA Toshiaki  Department of Life Science, Tokyo Institute of Technology, 生命理工学部, 教授 (10088859)

Co-Investigator(Kenkyū-buntansha) SHILLING Fraser  Department of Physiology, The University of Connecticut Health Center, コネチカット大学・医学部, 研究員
JAFFE Laurinda  Department of Physiology, The University of Connecticut Health Center, コネチカット大学・医学部, 教授
CHIBA Kazuyoshi  Department of Life Science, Tokyo Institute of Technology, 生命理工学部, 助手 (70222130)
NISHINA Hiroshi  Department of Life Science, Tokyo Institute of Technology, 生命理工学部, 助手 (60212122)
TAKAHASHI Katsunobu  Department of Life Science, Tokyo Institute of Technology, 生命理工学部, 助手 (40183850)
FRASER Shill  コネチカット大学, 医学部(米国), 研究員
LAURINDA A J  コネチカット大学, 医学部(米国), 教授
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1992: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1991: ¥3,500,000 (Direct Cost: ¥3,500,000)
KeywordsGTP-binding proteins / pertussis toxin / 1-methyladenine / Oocyte maturation / Fertilization / Membrae receptors
Research Abstract

In response to a meiosis-inducing hormone, 1-methyladenine (1-MA), starfish oocytes undergo reinitiation of meiosis with germinal vesicle breakdown (GVBD). The 1-MA-initiated GVBD is, however, inhibited by prior microinjection of pertussis toxin into the oocytes, suggesting that a pertussis toxin-sensitive GTP-binding protein (G protein) is involved in the 1-MA-induced signal transduction. Based on these findings, we purified a G protein serving as the substrate of pertussis toxin from the plasma membranes of starfish oocytes. The purified G protein had an alphabetagamma-trimeric structure consisting of 39-kDa alpha, 37-kDa beta, and 8-kDa gamma-subunits. The 39-kDa alpha subunit contained a site for ADP-ribosylation catalyzed by pertussis toxin. The purified starfish G protein had a GTP-binding activity with a high affinity and displayed a low GTPase activity, The activity of the G protein serving as the substrate for pertussis toxin-catalyzed ADP-ribosylation was inhibited by its ass … More ociation with a non-hydrolyzable GTP analogue. Thus, the starfish G protein appeared to be similar to mammalian G proteins at least in terms of its structure and properties of nucleotide binding and pertussis toxin substrate. The microinjection of the purified betagamma-subunits into the oocytes induced GVBD as had been observed with 1-MA. There were apparently two forms of 1-MA receptors with a high and a low affinities in the membranes. The high-affinity form was converted into the low-affinity one in the presence of a non-hydrolyzable analogue of GTP. The 39-kDa alpha-subunit of the starfish G protein was also ADP-ribosylated by cholera toxin only when 1-MA was added to the membranes. The ADP-ribosylated 39-kDa alpha-subunit could be immunoprecipitated with an antibodies raised against the carboxy-terminal site of mammalian inhibitory G-alpha. These results indicate that 1-MA receptors are functionally coupled with the pertussis toxin-substrate G protein and that the betagamma-complex resolved from alpha-subunit is an active molecule for the initiation of GVBD in starfish oocytes. Less

Report

(2 results)
  • 1992 Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] TADENUMA,Hirohiko(蓼沼 裕彦): "Purification and characterization of a GTP-binding protein serving as pertussis toxin substrate in starfish oocytes" Archieves of Biochemistry and Biophysics. 290. 411-417 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] CHIBA,Kazuyoshi(千葉 和義): "The primary structure of the α-subunit of a starfish G protein involved in 1-methyladenine-induced oocyte maturation" European Journal of Biochemistry. 207. 833-838 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] TADENUMA,Hirohiko(蓼沼 裕彦): "Properties of 1-methyladenine receptors in starfish oocyte membranes: Involvement of pertussis toxin-sensitive GTP-binding protein in the receptor-mediated signal transduction" Biochemical and Biophysical Research Communications. 186. 114-121 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] TAKEI,Yoshinori(武井 義則): "A GTP-binding protein in rat liver nuclei serving as the specific substrate of pertussis toxin-catalyzed ADP-ribosylation" Journal of Biological Chemistry. 267. 5085-5089 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] KONTANI,Kenji(紺谷 圏二): "Molecular heterogeneity of the βγ-subunits of GTP-binding proteins in bovine brain membranes" Archives of Biochemistry and Biophysics. 294. 527-533 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] KATADA,Toshiaki(堅田 利明) (eds.Hiroshi YOSIDA & Michi Ui): "Neurotransmitter Receptors and Intracellular Signaling" Excerpta Medida(Amsterdam), 91 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] KATADA,Toshiaki(堅田 利明) (eds.Hiroshi YOSIDA, & Hideaki KAWAGUCHI): "New Aspects in the treatment of Failing Heart" Springer-Verlag(Berlin), 241 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hirohiko Tadenuma, et al.: "Purification and characterization of a GTP-binding protein serving as pertussis toxin substrate in starfish oocytes" Archieves of Biochemistry and Biophysics. 290. 411-417 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kazuyoshi Chiba, et al.: "The primary structure of the alpha-subunit of a starfish G protein involved in 1-methyladenine-induced oocyte maturation" European Journal of Biochemistry. 207. 833-835 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hirohiko Tadenuma, et al.: "Properties of 1-methyladenine receptors in starfish oocyte membranes : Involvement of pertussis toxin-sensitive GTP-binding protein in the receptor-mediated signal transduction" Biochemical and Biophysical Research Communications. 186. 114-121 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Yoshinori Takei, et al.: "A GTP-binding protein in rat liver nuclei serving as the specific substrate of pertussis toxin-catalyzed ADP-ribosylation" Journal of Biological Chemistry. 267. 5085-5089 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kenji Kontani, et al.: "Molecular heterogeneity of the betagamma-subunits of GTP-binding proteins in bovine brain membranes" Archives of Biochemistry and Biophysics. 294. 527-533 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Toshiaki Katada, et al. (eds. Hiroshi Yosida & Michi Ui): Excerpta Medica (Amsterdam). Neurotransmitter Receptors and Intracellular Signaling, 30-43 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Toshiaki Katada, et al. (eds. Hisakazu Yasuda, & Hideaki Kawaguchi: Springer-Verlag (Berlin). New Aspects in the treatment of Failing Heart, 53-57 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hirohiko Tadenuma: "Purification and characterization of a GTP-binding protein serving as pertussis toxin substrate in starfish oocytes" Archieves of Biochemistry and Biophysics. 290. 411-417 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Iiri Taroh: "Functional modification by cholera-toxin-catalyzed ADP-ribosylation of a guanine-nucleotide-binding regulatory protein serving as the substrate of pertussis toxin" European Journal of Biochemistry. 202. 635-641 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Urara Tomita: "Direct activation of GTP-binding proteins by venom peptides that contain cationic clusters within their alpha-helical structures" Biochemical and Biophysical Research Communications. 178. 400-406 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kazuyoshi Chiba: "The primary structure of the α-subunit of a starfish G protein involved in 1-methyladenine-induced oocyte maturation" European Journal of Biochemistry.

    • Related Report
      1991 Annual Research Report
  • [Publications] 浦川 紀元,唐木 英明編: "カルシウムと情報伝達系" 文永堂出版, 1-352 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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