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Identification and characterization of a novel cyclic GMP/G-kinase substrate protein

Research Project

Project/Area Number 04454148
Research Category

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

Allocation TypeSingle-year Grants
Research Field 薬理学一般
Research InstitutionNiigata University

Principal Investigator

IMAI Shoichi  Niigata University School of Medicine Department of Pharmacology Professor, 医学部, 教授 (60013869)

Co-Investigator(Kenkyū-buntansha) YUTAKA Yoshida  Niigata University School of Medicine Department of Pharmacology Assistant Profe, 医学部, 講師 (40182795)
NAKAZAWA Mikio  Niigata University School of Medicine Department of Pharmacology Associate Profe, 医学部, 助教授 (80143759)
Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordscGMP / G-Kinase / Porcine aortic smooth muscle / Calmodulin / 240 kDa protein / Heparin / IP^4 / IP^3 receptor / Concanavalin A / 240 kDaタンパク質 / 血管平滑筋 / イオンチャネル / cyclic GMP
Research Abstract

The 240 kDa, cGMP-dependent protein kinase (G-kinase) substrate obtained from porcine aortic smooth muscle as a protein, whose phosphorylation was closely associated with stimulation of plasma membrance Ca^<2+>-pump ATPase(J.Biol.Chem., 266,19819-19825,1990), was purified to near homogeneity by three successive chromatographic runs with calmodulin -, concanavalin A - and heparin-Sepharose columns from Triton X-100 solubilzed microsomes. The purified protein was found to bind inositol 1,4,5-trisphosphate (IP^3)in a specific, heparin-inhibitable manner (Kd : 2.0nM ; Bmax : 450 pmol/mg protein). The protein also bound inositol 1,3,4,5, -tetrakisphosphate, though weakly. In sedimentation experiments on a linear sucrose density gradient the IP^3 binding activity was always with the 240-kDa protein. G-kinase phosphorylated the IP^3 receptor purified from rat cerebellum as well as the 240-kDa protein. Sialic acid content of the protein measured with Limulus polyphemus agglutinin was not significantly different from that of the cerebellar IP^3 receptor. Threr were, however, some differences : On SDS-PAGE the 240-kDa protein presented itself as two polypeptides with similar, but slightly differing Mr's both of which could be phosphlrylated by G-kinase, while cerebellar IP^3 receptor presented itself as a single band, and only the 240 kDa protein was found to bind with calmodulin. A certain immunological differences were also found. Thus, the 240 kDa protein may be a new member of IP^3 receptor superfamily.

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] D.-L.Luo,M.Nakazawa,T.Ishibashi,K.Kato,S.Imai: "Putative,selective inhibitors of sarcoplasmic reticulum Ca^<2+>-pump ATPase inhibit relaxation by nitroglycerin and atrial natriuretic factor of the rabbit aorta contracted by phenylephrine" The Journal of Pharmacology and Experimental Therapeutics. 265. 1187-1192 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Ishibashi,M.Hamaguchi,K.Kato,T.Kawada,H.Ohta,H.Sasage,S.Imai: "Relationship between myoglobin contents and increases in cyclic GMP produced by glyceryl trinitrate and nitric oxide in rabbit aorta,right atrium and papillary muscle" Naunyn-Schmiedeberg's Archives of Pharmacology. 347. 553-561 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] D.-L.Luo et al.: "Putative, selective inhititors of sarcoplasmic reticulum Ca^<32+>-pump ATPase inhibit relaxation by nitroglycerin and atrial natriuretic factor of the rabbit aorta contracted by phenylephrine" The Journal of Pharmacology and Experimental Therapeutics. 265(3). 1187-1192 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Ishibashi et al.: "Relationship between myoglobin contens and increases in cyclic GMP produced by glyceryl trinitrate and nitric oxide in rabbit aorta, right atrium and papillary muscle" Naunyn-Schmiedeberg's Archives of Pharmacology. 347(5). 553-561 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] D.-L.Luo,M.Nakazawa,T.Ishibashi,K.Kato,S.Imai: "Putative,selective inhititors of sarcoplasmic reticulum Ca^<2+>-pump ATPase inhibit relaxation by nitroglycerin and atrial natriuretic factor of the rabbit aorta contracted by phenylephrine" The Journal of Pharmacology and Experimental Therapeutics. 265. 1187-1192 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.Ishibashi,M.Hamaguchi,K.Kato,T.Kawada,H.Ohta,H.Sasage,S.Imai: "Relationship between myoglobin contens and increases in cyclic GMP produced by glyceryl trinitrate and nitric oxide in rabbit aorta,right atrium and papillary muscle" Naunyn-Schmiedeberg's Archives of Pharmacology. 347. 553-561 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yutaka Yoshida,Ji-Qun Cai and Shoichi Imai: "Plasma membrane Ca^<2+>-pump ATPase is not a substrate for cGMP-dependent protein kinase" Journal of Biochemistry. 111. 559-562 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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