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

新型环 GMP/G 激酶底物蛋白的鉴定和表征

基本信息

  • 批准号:
    04454148
  • 负责人:
  • 金额:
    $ 0.64万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
  • 财政年份:
    1992
  • 资助国家:
    日本
  • 起止时间:
    1992 至 1993
  • 项目状态:
    已结题

项目摘要

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.
从猪主动脉平滑肌获得的240 kDa cGMP依赖性蛋白激酶(G-激酶)底物作为蛋白质,其磷酸化与质膜Ca^2+泵ATP酶的刺激密切相关(J.Biol.Chem.,266,19819 - 19825,1990),通过三次连续的层析操作,用钙调蛋白-、伴刀豆球蛋白A -和肝素-琼脂糖柱从Triton X-100溶解的微粒体中纯化至接近均一。发现纯化的蛋白质以特异的肝素可降解的方式与1,4,5-三磷酸肌醇(IP ^[3])结合(Kd:2.0 nM; Bmax:450 pmol/mg蛋白质)。该蛋白也结合肌醇1,3,4,5-四磷酸,虽然很弱。在线性蔗糖密度梯度的沉降实验中,IP^3的结合活性总是与240 kDa的蛋白质结合。G激酶磷酸化了从大鼠小脑中纯化的IP^3受体以及240 kDa蛋白质。用鲎凝集素测得的蛋白质唾液酸含量与小脑IP^3受体的唾液酸含量没有显著差异。但也有一些不同之处:在SDS-PAGE上,240-kDa蛋白质呈现为两种多肽,其Mr相似但略有不同,这两种多肽都可以被G-激酶磷酸化,而小脑IP^3受体呈现为单一条带,并且只有240-kDa蛋白质与钙调素结合。结果表明,该蛋白可能是IP^3受体超家族的一个新成员。

项目成果

期刊论文数量(13)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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 (1
T.Ishibashi 等人:“兔主动脉、右心房和乳头肌中肌红蛋白浓度与三硝酸甘油和一氧化氮产生的环 GMP 增加之间的关系”Naunyn-Schmiedeberg 药理学档案。
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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
D.-L.Luo 等人:“推定的肌浆网 Ca^<32>-泵 ATP 酶选择性抑制剂可抑制去氧肾上腺素收缩的兔主动脉的硝酸甘油和心房钠尿因子的舒张”《药理学与实验杂志》
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    0
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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 A
T.Ishibashi,M.Hamaguchi,K.Kato,T.Kawada,H.Ohta,H.Sasage,S.Imai:“兔主动脉中肌红蛋白含量与三硝酸甘油酯和一氧化氮产生的环 GMP 增加之间的关系,右
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    0
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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 P
D.-L.Luo、M.Nakazawa、T.Ishibashi、K.Kato、S.Imai:“推定的选择性肌浆网 Ca^<2>-泵 ATP 酶抑制剂抑制兔硝酸甘油和心房利尿钠因子的松弛
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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
T.Ishibashi,M.Hamaguchi,K.Kato,T.Kawada,H.Ohta,H.Sasage,S.Imai:“兔主动脉中肌红蛋白含量与三硝酸甘油酯和一氧化氮产生的环 GMP 增加之间的关系,右
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IMAI Shoichi其他文献

IMAI Shoichi的其他文献

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{{ truncateString('IMAI Shoichi', 18)}}的其他基金

Effects of drugs on the ischemic changes in cardiac function, myocardial energy metabolism and ion permeability-A study by nuclear magnetic resonance spectroscopy (NMR)9
药物对缺血性心功能、心肌能量代谢及离子通透性变化的影响——核磁共振波谱(NMR)研究9
  • 批准号:
    60480126
  • 财政年份:
    1985
  • 资助金额:
    $ 0.64万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)

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