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Intracellular distribution and regulatory function of protein phosphatases

Research Project

Project/Area Number 09670042
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionNagoya University

Principal Investigator

TAKAI Akira  School of Medicine, Nagoya University Associate Professor, 医学部, 助教授 (50126869)

Co-Investigator(Kenkyū-buntansha) TOKUNO Hiroyuki  School of Medicine, Res.Assoc., 医学部, 助手 (60155520)
KUBA Kenji  School of Medicine, Professor, 医学部, 教授 (60080561)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
KeywordsProtein phosphatases / Protein phosphorylation / Signal transduction / Okadaic acid / Microcystins / Tautomycin / CRTR / Ciliary muscle / ト-トマイシン / 酵素阻害剤
Research Abstract

The aim of this project has been to investigate the roles of reversible protein phosphorylation in regulation of cell motility and ion transport. During the term of this project we carried out physiological and biochemical experiments chiefly in smooth and cardiac muscles, as summarized below. Part of the results obtained have been published (see the reprints attached).
(1) Okadaic acid (OA) is a potent inhibitor of protein phosphatase 1 and 2A (PP1 and PP2A) having a higher affinity to PP2A than to PP1. In smooth muscle preparations with intact plasma membrane, OA reversibly inhibits contraction and myosin light-chain (MLC) phosphorylation. Another PP inhibitor, tautomycin, which exhibits a higher affinity PP1 than PP2A is known to enhance contraction and myosin light chain phosphorylation. In the present experiments we have observed that OA strongly inhibits this contractile effect of tautomycin. The MLC phosphorylation was also decreased by addition of OA but about 10 % of MLC were s … More till phosphorylated even when contraction was completely inhibited by OA.These results indicate that there are several steps toward smooth muscle contraction which is suppressed by protein phosphorylation.
(2) In smooth muscle cells isolated from bovine ciliary body we have studied effects of carbachol on the membrane potential and current using the whole-cell clamp technique. We have demonstrated the existence of a non-selective cation channel which is activated by muscarinic receptors belonging to the M3 subtype. This channel, which admits Ca^<2+>, may serve at least partly as a Ca^<2+> entry required for sustained contraction.
(3) In isolated myocyte of guinea-pig ventricle, we have examined the effect of anthracene-9-carboxylic acid (9AC), a Cl^- channel inhibitor, on the CFTR channel. We have shown that the activating effect of isoprenaline or forskolin on the CFTR is reversibly enhanced and prolonged in the presence of 9AC.We have also shown that 9AC inhibits a fraction of intracellular p-nitrophenyl phosphatase activity, which is insensitive to known phosphatase inhibitors, such as OA, tartaric acid or bromotetramisole. Less

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] Takai,Y.,Awaya,S.& Takai,A.: "Activation of non-selective cation conductance by carbachol in freshly isolated bovine ciliary musclecells" Pflugers Archiv-European Journal of Physiology. 433(6). 705-712 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Zhou,S.S.,Takai,A.Tominaga,M.& Okada,Y.: "Phosphatase-mediated enhancement of cardiac cAMP-activated Cl^- conductance by a Cl^- channel blocker,anthracene-9-carboxylate" Circulation Research. 81. 219-228 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takemura,M.,Kitagawa,et al.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase α" Oncogene. 15. 2483-249〓 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yotsu-Yamashita,M.,et al.: "Effects of selective modifications of several hydroxyls of tetrodotoxin on its affinity to rat brain membrane" Journal of Pharmacology and Experimental Therapeutics. 印刷中. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 高井 章: "蛋白質脱燐酸化反応:解析の基本手技" 日本血栓止血学会誌. 8(6). 504-516 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 高井 章: "PP1およびPP2Aに特異的阻害作用を有する天然毒素" 蛋白質・核酸・酵素. 43(8). 1091-1101 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Botana,L.M.(ed.): "Seafood Toxicity: Mode of Action,Pharmacology and Physiology" Marcel Dekker,Inc.(New York)印刷中, (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] TAKAI,Y., AWAYA,S.& TAKAI,A.: "Activation of non-selective cation conductance by carbachol in freshly isolated bovine ciliary muscle cells." Pflugers Archiv-European Journal of Physiology. 433. 705-712 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] TAKEMURA,M., KITAGAWA,T., IZUTA,S., WASA,J., TAKAI,A., AKIYAMA,T.& YOSHIDA,S.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase alpha." Oncogene. 15. 2483-2492 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] ZHOU,S.S., TAKAI,A., TOMINAGA,M.&OKADA,Y.: "Phosphatase-mediated enhancement of cardiac cAMP-activated Cl^- conductance by a Cl^- channel blocker, anthracene-9-carboxylate." Circulation Research. 81. 219-228 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] TAKAI,A.& HARADA,K.I.: "Freshwater hepatotoxins : ecobiology and classification." In Sea Food Toxicity : Mode of Action, Pharmacology and Physiology of Phycotoxins, ed.BOTANA,L.M., New York : Marcel Decker. (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] TAKAI,A.: "Naturally occurring toxins with specific inhibitory activity against protein serine/threonine phosphatases 1 and 2A [Review] [Japanese]" Tanpakushitsu Kakusan Koso-Protein Nucleic Acid, Enzyme. 43. 1091-1101 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] YOTSU-YAMASHITA,M., SUGIMOTO,A., TAKAI,A.& YASUMOTO,T.: "Effects of specific modifications of several hydroxyls of tetrodotoxin on its affinity to rat brain membrane." Journal of Pharmacology & Experimental Therapeutics in the Press. (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takai,Y., Awaya,S.& Takai,A.: "Activation of non-selective cation conductance by carbachol in freshly isolated bovine ciliary muscle cells" Pflugers Archiv-European Journal of Physiology. 433 (6). 705-712 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Zhou,S.S., Takai,A.Tominaga,M.& Okada,Y.: "Phosphatase-mediated enhancement of cardiac cAMP-activated Cl^- conductance by a Cl^- channel blocker, anthracene-9-carboxylate" Circulation Research. 81. 219-228 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takemura,M, Kitagawa, et al.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase α" Oncogene. 15. 2483-2492 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yotsu-Yamashita,M., et al.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase α" Journal of Pharmacology and Experimental Therapeutics. 印刷中. (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] 高井 章: "蛋白質脱燐酸化反応:解析の基本手技" 日本血栓止血学会誌. 8 (6). 504-516 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] 高井 章: "PP1およびPP2Aに特異的阻害作用を有する天然毒素" 蛋白質・核酸・酸素. 43 (8). 1091-1101 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Botana,L.M.(ed.): "Seafood Toxicity : Mode of Action, Pharmacology and Physiology" Marcel Dekker, Inc. (New York) 印刷中, (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takai, Y., Awaya,S. & Takai, A.: "Activation of non-selective cation conductance by carbachol in freshly isolated bovine ciliary muscle cells" Pflugers Archiv-European Journal of Physiology. 433(6). 705-712 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Zhou,S.S., Takai,A. Tominaga,M. & Okada,Y.: "Phosphatase-mediated enhancement of cardiac cAMP-activated Cl^-conductance by a Cl^-channel blocker,anthracene‐9‐carboxylate" Circulation Research. 81. 219-228 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Takemura,M.., Kitagawa, et al.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase α" Oncogene. 15. 2483-2492 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 高井 章: "蛋白質脱燐酸化反応:解析の基本手技" 日本血栓止血学会誌. 8(6). 504-516 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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