Dynamisms and funstions of calpain in early embryogenesis

钙蛋白酶在早期胚胎发生中的动态和功能

• 批准号: 07458157
• 负责人: EMORI Yasufumi
• 金额: $ 5.18万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07458157/
• 关键词: calpain; calcium; Drosophila; signalling; cytoskeleton; actin-cytoskeleton; proteclysis; early embryogenesis; プロテアーゼ; 発生; 情報伝達

Changes in shapes of cells during cell division and cell differentiation are mainly dependent on actin-related cytoskeletons on the cell surface. Calpain, calcium-dependent neutral protease, has proteolytic activity which irreveribly changes the components of the cytoskeleton. In this study, the physiological functions of calpain have been analyzed by developmental and biochmical methods during early embryogenesis of Drosophila melanogaster. First, localization of calpain and actin was analyzed by immunohistochmical methods. Second, cDNA clones for actin-binding proteins, probable substrates for calpain, were obtained and expressed in Escherichia coli together with the expression of active calpain. Third, the probes for calpain reaction, specific antibodies for cleavage sites generated by calpain reaction, were prepared. Fourth, using these probes described above, dynamism of localization of calpain and its substrates were analyzed during Drosophila development. Consequently, the results were obtained that calpain is involved in the formation and reorganization of actin-related cytoskeleton in pseudo-cleavage furrow.

细胞分裂和分化过程中细胞形状的变化主要依赖于细胞表面与肌动蛋白相关的细胞骨架。钙蛋白酶是一种依赖钙的中性蛋白酶，具有不可逆地改变细胞骨架成分的蛋白水解活性。本研究采用发育和生化方法分析了黑腹果蝇早期胚胎发育过程中钙蛋白酶的生理功能。首先用免疫组化方法分析calpain和actin的定位。其次，获得了肌动蛋白结合蛋白（可能是calpain的底物）的cDNA克隆，并在大肠杆菌中与活性calpain的表达一起表达。第三，制备calpain反应探针，即calpain反应产生的裂解位点特异性抗体。第四，利用上述探针，分析了果蝇发育过程中钙蛋白酶及其底物的定位动态。结果表明，钙蛋白酶参与了假卵裂沟中肌动蛋白相关细胞骨架的形成和重组。

项目成果

A.S.Arora,P.D.Goen,Y.Emori & G.J.Gores: "A cascade of degradative hydrolase acting contributes to hepatocyte recrosis during anoxia" American Journal of Physiology. 270. G238-G245 (1996)

A.S.阿罗拉、P.D.Goen、Y.Emori

Y.Emori: "Receptors and intracellular signalling pathways (in Japanese)" Molecular Biology of Hormones. 137-152 (1996)

Y.Emori：“受体和细胞内信号传导途径（日语）”激素分子生物学。