Limited Proteolysis of Filamin Is Catalyzed by Caspase-3 in U937 and Jurkat Cells.

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概要

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• Members of the caspase family have been implicated as key mediators of apoptosis in mammalian cells. However, few of their substrates are known to have physiological significance in the apoptotic process. We focused our screening for caspase substrates on cytoskeletal proteins We found that an actin binding protein, filamin, was cleaved from280 kDa to 170, 150, and 120 kDa major N-terminal fragments, and 135, 120, and 110 kDa major C-terminal fragments when apoptosis was induced by etoposide in both the human monoblastic leukemia cell line U937, and the human T lymphoblastic cell line Jur-kat. The cleavage of filamin was blocked by a cell permeable inhibitor of caspase-3-likeprotease, Ac-DEVD-cho, but not by an inhibitor of caspase-1-like protease, Ac-YVAD-cho. These results suggest that filamin is cleaved by a caspase-3-like protease. To examinewhether caspase-3 cleaves filamin in vitro, we prepared a recombinant active form ofcaspase-3 directly using a Pichia pastoris overexpression system. When we applied recombinant active caspase-3 to the cell lysate of U937 and Jurkat cells, filamin wascleaved into the same fragments seen in apoptosis-induced cells in vivo. Platelet filamin was also cleaved directly from 280 kDa to 170, 150, and 120 kDa N-terminal fragments, and the cleavage pattern was the same as observed in apoptotic human cells in vivo. These results suggest that filamin is an in vivo substrate of caspase-3.

• 社団法人 日本生化学会の論文

著者

• Sasagawa Noboru

  Department Of Agricultural Chemistry The University Of Tokyo

• Ishiura Shoichi

  Department Of Life And Cognitive Sciences College Of Arts And Sciences The University Of Tokyo

• KAWAHARA Hiroyuki

  Institute of Molecular and Cellular Biosciences, The University of Tokyo

• SUZUKI Koichi

  Institute of Molecular and Cellular Biosciences, The University of Tokyo

• Kouchi Zen

  Institute Of Molecular And Cellular Biosciences The University Of Tokyo

• TOMIOKA Shigeo

  Institute of Applied Microbiology, University of Tokyo

• MAEDA Tatsuya

  Institute of Molecular and Cellular Bioscences, The University of Tokyo

• Umeda Tatsuya

  Institute of Moleccluar and Cellular Bioscienees, The Unitwrsityof Tokyo

• Sasagawa Noboru

  Department of Life Scencens., Graduate School Arts and Sciences, The Uniuersity of Tokyo

• Sorimach Hiroyuki

  Institute of Moleccluar and Cellular Bioscienees, The Unitwrsityof Tokyo

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