Developmental and molecular genetical analyzes on calpain-calpastatin system at individual and cellular levels

• Project/Area Number: 04454590
• Research Category: Grant-in-Aid for General Scientific Research (B)
• Allocation Type: Single-year Grants
• Research Field: 物質生物化学
• Research Institution: THE UNIVERSITY OF TOKYO
• Principal Investigator: EMORI Yasufumi The University of Tokyo, Graduate School of Science Associate Professor, 大学院・理学系研究科, 助教授 (60160389)
• Project Period (FY): 1992 – 1993
• Project Status: Completed (Fiscal Year 1993)
• Budget Amount: ¥6,200,000 (Direct Cost: ¥6,200,000); Fiscal Year 1993: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 1992: ¥4,600,000 (Direct Cost: ¥4,600,000)
• Keywords: Calpain / Calpastatin / Protease / Calcium / Signal transduction / Drosophila / Cytoskeleton / Melecula biology

Research Abstract

Calpain is an intracellular cysteine protease, possibly involved in Ca^<2+>-mediated signalling and activated by enhancement of intracellular Ca^<2+> concentration.
First, I have identified two Drosophila genes encoding calpains. Their mRNA expression was studied by Northern blotting analysis and in situ hybridization experiment, which revealed that the two Drosophila calpains belong to conventional calpain family showing ubiquitous distribution.
Next, I prepared a specific antibody against one type of the calpains. Using this antibody, cellular and subcellular localization of the calpain molecule was investigated in the early embryo. The results showed that calpain is localized with actin-related cytoskeletal structure in the very early embryo before cellularization, and in the precleavage furrows just before cellularization. These observations indicate that the Drosophila calpain is involved in univeral cellular functions, especially in the reorganization of actin-related cytoskeleton.
I also studied mode of interaction between calpain and calpastatin, an endogenous inhibitor specific for calpain, by biochemical methods, which provided useful informations for further molecular studies on in vivo functions of the Drosophila calpain-calpastatin system.

Research Products

• [Publications] 榎森康文・大海忍・牧正敏: "カルパインとカルパスタチン-構造と機能" 生化学. 64. 1201-1224 (1992)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] H.Kawasaki,Y.Emori & K.Suzuki: "Calpastatin has two distinct sites for interaction with Calpain-Effect of calpastatin frogments on the binding of calpain to membranes" Archives of Biochemistry and Biophysics. 305. 467-472 (1993)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 榎森康文: "タンパク質工学-基礎と応用-(カルシウムプロテアーゼとそのインヒビター)" 丸善株式会社 12ページ(総236ページ中), (1992)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Yasufumi Emori, Shinobu Ohmi and Masatosi Maki: "Calpain and Calpastatin ; Structure and Function." Seikagaku. 64. 1201-1224 (1992)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hiroshi Kawasaki, Yasufumi Emori and Koichi Suzuki: "Calpastatin Has Two Distinct Sites for Interaction with Calpain-Effect of Calpastatin Fragments on the Binding of Calpain to Membrabes." Archives of Biochemistry and Biophysics. 305. 467-472 (1993)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Yasufumi Emori: Calcium Protease and Its Inhibitor in Protein Engineering (K.Suzuki, ed.). Maruzen, 191-122 (1992)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] H.Kawasaki,Y.Emori & K.Suzuki: "Calpastatin has two distinct sites for interaction with Calpain-Effect of calpastatin fragments on the binding of calpain to membranes." Archives of Biochemistry and Biophysics. 305. 467-472 (1993)
• [Publications] 榎森 康文・大海 忍・牧 正敏: "カルパインとカルパスタチンー構造と機能ー" 生化学. 64. 1201-1224 (1992)
• [Publications] 伊藤 菁茂,稲垣 冬彦,榎森 康文,鏡山 博行,菊池 正和,熊谷 泉,神田 大輔,高橋 信弘,綱沢 進,西山 真,林 秀行,別府 輝彦,松井 裕,松沢 洋,村上 和雄,矢内原 昇,油谷 克美: "タンパク質工学ー基礎と応用ー" 丸善株式会社, 236 (1992)