Molecularphysiological analysis of calcium-signaling proteins in cardiac sarcoplasmic reticulum

心脏肌浆网钙信号蛋白的分子生理学分析

• 批准号: 02404044
• 负责人: TADA Michihiko
• 金额: $ 14.46万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02404044/
• 关键词: Isolated Cardiomycyte; Intracelluar Ca^<2+> Content Measuring System; cardiac SR; Intracellular Ca Transient; phospholamban; Synthetic Peptide; Ca^<2+> Pump ATPase; Monoclonal Antibody

We investigated the molecular mechanism of regulation of the proteins of cardiac sarcoplasinic reticulum (SR) which has pivotal role in excitation-contraction coupling of myocardium.1. Physiological roles of Ca signaling in regulation of cardiac property : We investigated the effects of a monoclonal antibody against phospholamban, one of the Ca signaling proteins in cardiac SR, on ATPase activity of cardiac SR Ca pump ATPase. We found that the antibody inhibit the direct protein-protein interaction between the two SR proteins (J. Mol. Cell. Cardiol. 23, 1223, 1991). We investigated the effects of synthetic phospholamban peptides on the ATPase. The dual regulatory effects of pliospholainban on Ca pump ATPase were revealed ; the cytoplasmic domain for V_<max> and the intramembrane domain for K_<Ca> (J. Biol. Chem. 267, 1647, 1992).2. Pathological roles of Ca regulatory system of cardiac SR in normal and stressed inyocytes : We established the intracellular Ca^<2+> content measuring system in isolated cardiac myocyte, and evaluated intracellular Ca^<2+> transients of cardiac myocyte under hypoxia and Ca overload. Currently, we are on the way to evaluate the effect of anti-phospholamban monoclonal antibody and the synthetic peptides on the Ca^<2+> transient. We are examining the regulation of the expression of cardiac SR Ca signaling proteins in normal, stressed, or cardioinyopatliic cardiomyocyte.

我们研究了在心肌兴奋-收缩偶联中起关键作用的心肌肌浆网（sarcoplasinic reticulum，SR）蛋白的调控分子机制. Ca信号在心脏特性调节中的生理作用：我们研究了抗受磷蛋白（心脏SR中的Ca信号蛋白之一）的单克隆抗体对心脏SR Ca泵ATP酶活性的影响。我们发现抗体抑制两种SR蛋白之间的直接蛋白质-蛋白质相互作用（J. Mol. Cell. Cardiol. 23，1223，1991）。我们研究了合成受磷蛋白肽对ATP酶的影响。发现pliospholainban对Ca泵ATP酶的双重调节作用：胞质结构域对V_<max>和膜内结构域对K_<Ca>（J.Biol.Chem.267，1647，1992）.心肌SR钙调节系统在正常和应激心肌细胞中的病理作用：建立了心肌细胞内Ca^&lt;2+&gt;含量测定系统，观察了缺氧和钙超载条件下心肌细胞内Ca^&lt;2+&gt;瞬变。目前，我们正在评估抗受磷蛋白单克隆抗体和合成肽对Ca^&lt;2+&gt;瞬变的影响。我们正在研究在正常、应激或心源性心肌细胞中心脏SR Ca信号蛋白表达的调节。

项目成果

Fujii, J.: "Co-expression of slow/cardiac muscle Ca^<2+>-ATPase (SERCA2) and phospholamban." FEBS Lett.273. 232-234 (1990)

Fujii, J.：“慢肌/心肌Ca^2-ATP酶(SERCA2)和受磷蛋白的共表达。”

Kimura,Y.: "Effects of monoclonal antibody against phospholamban on calcium pump ATPase of cardiac sarcoplasmic reticulum." J.Mol.Cell.Cardiol.23. 1223 (1991)

Kimura,Y.：“抗磷蛋白单克隆抗体对心脏肌浆网钙泵 ATP 酶的影响。”

Kuzuya,T.: "Detection of oxygenーderived free radical generation in the canine postischemic heart during late phase of reperfusion." Circ.Res.66. 1160-1165 (1990)

Kuzuya, T.：“再灌注后期犬缺血后心脏中氧自由基生成的检测。”Circ.Res.66 (1990)。

Inui,M.et al.: "Molecular mechanism of calcium uptake and release by cardiac sarcoplasmic reticulum" Jpn.Circ J.54. 1185-1191 (1990)

Inui,M.et al.：“心脏肌浆网吸收和释放钙的分子机制”Jpn.Circ J.54。

Fujii, J.: "Structure of the rabbit phospholamban gene, cloning of the human cDNA, and assignment of the gene to human chromosome 6." J. Biol. Chem.266. 11669-11675 (1991)

Fujii, J.：“兔受磷蛋白基因的结构、人类 cDNA 的克隆以及将该基因分配给人类 6 号染色体。”