Calpain-dependent proteolytic cleavage of the p35 cyclin-dependent kinase 5 activator to p25

被引:334
作者
Kusakawa, G
Saito, T
Onuki, R
Ishiguro, K
Kishimoto, T
Hisanaga, S
机构
[1] Tokyo Metropolitan Univ, Grad Sch Sci, Dept Biol Sci, Tokyo 1920397, Japan
[2] Tokyo Inst Technol, Fac Biosci, Lab Cell & Dev Biol, Midori Ku, Yokohama, Kanagawa 2268501, Japan
[3] Japan Womens Univ, Dept Chem & Biol Sci, Bunkyo Ku, Tokyo 1128681, Japan
[4] Mitsubishi Kasei Inst Life Sci, Tokyo 1948511, Japan
关键词
D O I
10.1074/jbc.M907757199
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cyclin-dependent kinase 5 (CDK5) is a unique CDK, the activity of which can be detected in postmitotic neurons. To date, CDK5 purified from mammalian brains has always been associated with a truncated form of the 35-kDa major brain specific activator (p35, also known as nck5a) of CDK5, known as p25. In this study,we report that p35 can be cleaved to p25 both in nitro and in vivo by calpain. In a rat brain extract, p35 was cleaved to p25 by incubation with Ca2+. This cleavage was inhibited by a calpain inhibitor peptide derived from calpastatin and was ablated by separating the p35.CDK5 from calpain by centrifugation. The p35 recovered in the pellet after centrifugation could then be cleaved to p25 by purified calpain. Cleavage of p35 was also induced in primary cultured neurons by treatment with a Ca2+ ionophore and Ca2+ and inhibited by calpain inhibitor I. The cleavage changed the solubility of the CDK5 active complex from the particulate fraction to the soluble fraction but did not affect the histone H1 kinase activity. Increased cleavage was detected in cultured neurons undergoing cell death, suggesting a role of the cleavage in neuronal cell death.
引用
收藏
页码:17166 / 17172
页数:7
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