Suppression of calpain-dependent cleavage of the CDK5 activator p35 to p25 by site-specific phosphorylation

Hirotsugu Kamei, Taro Saito, Mirai Ozawa, Yuichi Fujita, Akiko Asada, James A. Bibb, Takaomi C. Saido, Hiroyuki Sorimachi, Shin Ichi Hisanaga

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

Cdk5 is a proline-directed Ser/Thr protein kinase predominantly expressed in postmitotic neurons together with its activator, p35. N-terminal truncation of p35 to p25 by calpain results in deregulation of Cdk5 and contributes to neuronal cell death associated with several neurodegenerative diseases. Previously we reported that p35 occurred as a phosphoprotein, phospho-p35 levels changed with neuronal maturation, and that phosphorylation of p35 affected its vulnerability to calpain cleavage. Here, we identify the p35 residues Ser 8 and Thr138 as the major sites of phosphorylation by Cdk5. Mutagenesis of these sites to unphosphorylatable Ala increased susceptibility to calpain in cultured cells and neurons while changing them to phosphomimetic glutamate-attenuated cleavage. Furthermore, phosphorylation state-specific antibodies to these sites revealed that Thr138 was dephosphorylated in adult rat, although both Ser8 and Thr 138 were phosphorylated in prenatal brains. In cultured neurons, inhibition of protein phosphatases converted phosho-Ser8 p35 to dual phospho-Ser8/Thr138 p35 and conferred resistance to calpain cleavage. These results suggest phosphorylation of Thr138 predominantly defines the susceptibility of p35 to calpain-dependent cleavage and that dephosphorylation of this site is a critical determinant of Cdk5-p25-induced cell death associated with neurodegeneration.

Original languageEnglish (US)
Pages (from-to)1687-1694
Number of pages8
JournalJournal of Biological Chemistry
Volume282
Issue number3
DOIs
StatePublished - Jan 19 2007

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Kamei, H., Saito, T., Ozawa, M., Fujita, Y., Asada, A., Bibb, J. A., Saido, T. C., Sorimachi, H., & Hisanaga, S. I. (2007). Suppression of calpain-dependent cleavage of the CDK5 activator p35 to p25 by site-specific phosphorylation. Journal of Biological Chemistry, 282(3), 1687-1694. https://doi.org/10.1074/jbc.M610541200