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Studies on Physiological Roles of Protein Phosphatases in Mammalian Smooth Muscle Tissues

Research Project

Project/Area Number 01570062
Research Category

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

Allocation TypeSingle-year Grants
Research Field Neurophysiology and muscle physiology
Research InstitutionNagoya University

Principal Investigator

TAKAI Akira  Nagoya Univ., Physiology, Assoc. Professor, 医学部, 助教授 (50126869)

Co-Investigator(Kenkyū-buntansha) NAKAYAMA S.  Nagoya Univ. Physiology, Res. Associate, 医学部, 助手 (30192230)
TOKUNO H.  Nagoya Univ., Physiology, Res. Associate, 医学部, 助手 (60155520)
TOMITA T.  Nagoya Univ., Physiology, Professor, 医学部, 教授 (50078763)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1989: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsOkadaic acid / Protein phosphatase / Protein phosphorylation / Smooth muscle / Ionic channel / P-Nitrophenyl phosphate / pーニトロフェニン燐酸 / 蛋白質、燐酸化 / 気管枝喘息 / イソプレナリン / カルシウム依存性カリウムチャネル / パッチクランプ方
Research Abstract

In mammalian smooth muscle tissues, the composition and physiological functions of protein phosphatases have been studied with the use of okadaic acid (OA), a potent inhibitor of type 2A and type 1 protein phosphatases, and inhibitor 2, an intrinsic inhibitory factor of type 1 phosphatase. We have obtained the following results :
[1] In the extracts of smooth muscle tissues of guinea-pig, we have identified the following four distinct protein phosphatase activities : an inhibitor 2-sensitive (type 1) phosphatase activity which is susceptible to OA, a Mg^<2+>-dependent and OA-insensitive (type 2C) phosphatase activity, and two type 2A-like phosphatase activities with different susceptibilities to OA. We have also obtained results indicating that the type 2A-like fraction with lower affinity to okadaic acid may be responsible for dephosphorylation of myosin light chains to induce relaxation of smooth muscle.
[2] In isolated myocytes of rabbit trachea, we have shown that the openstate probability of Ca^<2+>-dependent K^+-channel is reversibly increased by either extracellular application of isoprenaline or intracellular application of protein kinase A. We have also observed that this effect is significantly enhanced and prolonged in the presence of okadaic acid.
[3] So far the dephosphorylation of p-nitrophenyl phosphate (pNPP) by protein phosphatase preparations has been thought to be due to contamination by alkaline phosphatases. We have found that the activity of protein phosphatases are very susceptible to OA, to the same extent as is their activity against phosphorylated proteins. As OA has been shown to have no effect on alkaline phosphatases, this result indicates that the activity against pNPP is intrinsic to the protein phosphatases.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] TAKAI,A.,TROSCHKA,M.,MIOSKES,G.& SOMLYO,A.V.: "Protein phosphatase composition in the smooth muscle of guineapig ileum studied with okadaic acid and inhibitor 2" Biochemical Journal(London). 262. 617-623 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] KUME,H.,TAKAI,A.,TOKUNO,H.& TOMITA.T.: "Regulation of Ca^<2+>-dependent K^+channel activity in tracheal myocytes by phosphorylation" Nature (London). 341. 152-154 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] OBARA,K.,TAKAI,T.,Ruegg,J.C.& de Lanerolle,P: "Okadaic acid,a phosphatase inhibitor,produces a Ca^<2+>-and calmodulinーindependent contraction of smooth muscle" Pfliigers Archiv. 414. 134-138 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 高井 章: "海綿毒素オカダ酸による蛋白質脱燐酸化過程の阻害と筋収縮増強作用。" 血管. 12. 61-66 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 高井 章: "Okadaic acidのプロテインホスファタ-ゼ抑制作用とその応用" 実験医学. 8. 1220-1226 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] TAKAI,A.& MIESKES,G: "Inhibitory effect of okadaic acid on the pーnitrophenylーphosphate phosphatase activity of protein phosphatases" Biochemical Journal (London).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 高井 章: "海綿毒素オカダ酸の蛋白質脱燐酸化酵素阻害作用ー高親和性阻害剤としての反応速度論的特性ー" 生物物理. 31. 13-17 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 高井 章: "オカダ酸の蛋白質脱燐酸化酵素阻害作用ーその特徴と応用ー" 神経研究の進歩.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Takai, A., Troschka, M., Mieskes, G. & Somlyo, A. V.: "Protein phosphatase composition in the smooth muscle of guineaーpig ileum studied with okadaic acid and inhibitor 2" Biochem. J.262. 617-623 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kume, H., Takai, A., Tokuno, H. & Tomita, T.: "Regulation of Ca^<2+>-dependent K^+-channel activity in tracheal myocytes by phosphorylation" Nature. 341. 152-154 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Takai, A. & Mieskes, G.: "Inhibitory effect of okadaic acid on the pーnitropheny phosphate phosphatase activity of protein phosphatases" Biochem. J.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] TAKAI,A.,TROSCHKA,M.,Mioskes,G.& SOMLYO,A.V.: "Protein phosphatase composition in the smooth muscle of guineapig ileum studied with okadaicacid and inhibitor 2" Biochemical Journal(London). 262. 617-623 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] KUME,H.,TAKAI,A.,TOKUNO,H.& TOMITA.T.: "Regulation of Ca^<2+>ーdependent K^+channel activity in tracheal myocytes by phosphorylation" Nature(London). 341. 152-154 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] OBARA,K.,TAKAI.T.,Ru^^″egg,J.C.& de Lanerolle,P.: "Okaclaic acid,a phosphatase inhibitor,produces a Ca^<2+>ーand calmodulinーindependent contraction of smooth muscle" Pflu^^″gers Archiv. 414. 134-138 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 高井 章: "海綿毒素オカダ酸による蛋白質・脱燐酸化過程の阻害と筋収縮増強作用" 血管. 12. 61-66 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 高井 章: "Okadaic acidのプロテインホナファ-ゼ抑制作用とその応用" 実験医学. 8. 1220-1226 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] TAKAI,A.& Mieskes,G: "Inhibitory effect of okadaic acid on the pーnitrophenylーphosphate phosphatase activity of protein phosphatases" Biochemical Journal(London).

    • Related Report
      1990 Annual Research Report
  • [Publications] 高井 章: "海綿毒素オカダ酸の蛋白質脱燐酸化酵素阻害作用ー高親和性阻害剤としての反応速度論的特性ー" 生物物理. 31. 13-17 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 高井 章: "オカダ酸の蛋白質脱燐酸化酵素阻害作用ーその特徴とその応用ー" 神経研究の進歩.

    • Related Report
      1990 Annual Research Report
  • [Publications] TAKAI,A.,Troschka,M.,Mieskes,G.,& Somlyo,A.V.: "Protein phosphatase composition in the smooth muscle of guinea-pig ileum studied with okudaic acid and inhibitor 2" Biochenicul Journal. 262. 617-623 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Kume,H.,TATAI,A.,TOKUNO,H.,& TO MITA,T.: "Regulation of Ca^<2+> dependent K^+ channel activity in tracheal myocytes by phosphorylation" Nature. 341. 152-154 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] OBARA,K.,TAKAI,A.,RUegg,J.C.& de Kaberikkem P: "Okadaic acid,a phosphatase inhibitor, produces a Ca^<2+> and calmodulin-inclependent contraction of smooth muscle" Pflugers Archiv. 414. 134-138 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 高井章: "海綿毒素オカダ酸による蛋白質脱燐酸化過程の阻害と筋収縮増強作用" 血管. 12. 61-66 (1989)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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