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1988 Fiscal Year Final Research Report Summary

Study of the mechanism of phospholipase C activation via a new GTP binding protein

Research Project

Project/Area Number 62571026
Research Category

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

Allocation TypeSingle-year Grants
Research Field 応用薬理学・医療系薬学
Research InstitutionFukushima Medical College

Principal Investigator

NAKAHATA Norimichi  Associtate Professor, Department of Pharmacology, Fukushima Medical College, 医学部薬理学講座, 助教授 (60045804)

Project Period (FY) 1987 – 1988
Keywordspertussis toxin (IAP) / phospholipase C / GTP binding protein / フォトアフィニテイーラベリング / 脱感作
Research Abstract

This study was undertaken to elucidate the GTP binding protein to activate phospholipase C (PLase C) which is not a substarate for pertussis toxin (IAP) in human astrocytoma cells. The stimulation of TXA_2-receptors as well as muscarinic, H_1-histamine and bradykinin resulted in activation of PLase C in an IAP insensitive manner. The GTP binding protein involved in these agonists-induced PLase C activations is not a substrate for IAP. Pretreatment of cells with agonists elicited the reduction of GTPgammaS-induced accumulation of inositol phosphates (IP) in membrane preparations, reflecting from a functional reduction of signal transduction after agonist treatment of intact cells. Heavy peak fraction of membranes after sucrose gradient separation contained receptors and GTP binding proteins. Receptors together with GTP binding proteins moved from heavy membrane fraction to light peak fraction after treatment of the intact cells with agonists. The reduction of GTP binding protein in heav … More y peak fraction after agonist treatment was analyzed by a photoaffinity labelling with [^<35>S]GTPgammaS. The 32 kDa GTP binding protein was reduced in Heavy peak fraction after treatments of the intact cells with agonists. The 32 kDa GTP binding protein might be involved in receptor-mediated activation of phospholipase C. Then, the 32 kDa GTP binding protein was purified from the porcine brain membranes. There is a 32 kDa GTP binding protein in the membranes, determined by a photoaffinity labelling with [^<32>P]alpha-GTP. The 32 kDa GTP binding protein was solubilized with 1 % lubrol from cholate-inextracted membranes. The 32 kDa gtp binding protein might be hydrophobic, and was not ADP-ribosylated with IAP. The 32 kDa GTP binding protein was purified by column chromatographies of DEAE-sephacel, sephacryl S-200 and hydroxyapatite. Although the 32 kda protein corresponded to [^<35>S]GTPgammaS binding activities was appeared in hydroxyapatite column chromatography, further purifications were necessary. In conclusion, there is the 32 kDa GTP binding protein in brain membranes as well as astrocytoma cells which is a candidate for a GTP binding pritein to activate phospholipase C in an IAP-insenitive manner. Less

  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] Norimichi Nakahata.;Hironori Nakanishi: J.Pharmacol.Exp.Ther.246. 635-640 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Norimichi Nakahata,et al.: Eur.J.Pharmacol.(1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Norimichi Nakahata,et al.: Biochem.Biophys.Res.Cummun.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Norimichi Nakahata; Hironori Nakanishi: "Bradykinin-induced contraction was inhibited by tiaramid, an anti-inflammatory drug, with an inhibition of increase in intracellular free calcium" J. Pharmacol. Exp. Ther.246. 635-640 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Norimichi Nakahata; Isao Matsuoka; Tomoyuki Ono; Hironori Nakanishi: "Thromboxane A_2 activates phospholipase C in astrocytoma cells via pertussis toxin-insensitive G-protein" Eur. J. Pharmacol.(1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Norimichi Nakahata; Isao Matsuoka; Takashi Suzuki; Hironori Nakanishi: "GTPgammaS binding activities were reduced in heavy membrane fraction during desensitization by Ca-mobilizing agonists in human astrocytoma cells" Biochem. Biophy. Res. Cummun.(1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Norimichi Nakahata; Isao Matsuoka; Takashi Suzuki; Hironori Nakanishi: "Solubilization of 32 kDa GTP binding protein from rabbit and porcine brain membranes"

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

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Published: 1990-03-20  

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