Proteolytic processing of nuclear factor κB by calpain in vitro

FEBS Letters

Sasaki, Makoto¹  Kunimatsu, Mitoshi¹  Liu, Zhen-Qiu²  Yang, Jian-Ping²  Ozaki, Yasuhiko¹  Okamoto, Takashi² 

[1] Department of Biochemistry, Nagoya City University Medical School, Mizuho-ku, Nagoya 467, Japan;Department of Molecular Genetics, Nagoya City University Medical School, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467, Japan

关键词: Nuclear factor κB;    DNA binding;    EMSA;    Calpain;    Protease inhibitor;    Western blot;    Gene expression;    NF-κB;    nuclear factor κB;    EMSA;    electrophoretic mobility shift assay;    PMSF;    phenylmethanesulfonyl fluoride;    EDTA;    ethylenediaminetetraacetic acid;    EGTA;    ethyleneglycol bis (2-aminoethyl-ether) tetraacetic acid;   

DOI  :  10.1016/0014-5793(96)00360-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Nuclear factor KB (NF-κB) is a transcription factor that is critical for the inducible expression of multiple cellular and viral genes. Using the electrophoretic mobility shift assay, we demonstrated that DNA binding activity of NF-κB was abolished by proteolysis with μ- and m-calpains in vitro. The proteolysis of NF-κB by calpains and hence the abolition of its DNA binding was prevented by calpastatin, calpain inhibitor I and proteasome inhibitor. We also provided evidence that calpains degrade the Cterminal domain of NF-κB by Western blot using anti-NF-κB (p65) C-terminal antibody. These observations indicate that calpains regulate gene expression through processing of NF-κB.

【 授权许可】

Unknown