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1993 Fiscal Year Final Research Report Summary

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
KeywordscGMP / G-Kinase / Porcine aortic smooth muscle / Calmodulin / 240 kDa protein / Heparin / IP^4 / IP^3 receptor / Concanavalin A
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.

  • Research Products

    (4 results)

All Other

All Publications (4 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
  • [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
  • [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
  • [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

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Published: 1995-03-27  

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