Proteolytic activation of protein kinase C δ by an ICE-like protease in apoptotic cells

Y. Emoto, Y. Manome, G. Meinhardt, H. Kisaki, S. Kharbanda, M. Robertson, T. Ghayur, W. W. Wong, R. Kamen, R. Weichselbaum, D. Kufe

Research output: Contribution to journalArticle

636 Scopus citations

Abstract

These studies demonstrate that treatment of human U-937 cells with ionizing radiation (IR) is associated with activation of a cytoplasmic myelin basic protein (MBP) kinase. Characterization of the kinase by gel filtration and in-gel kinase assays support activation of a 40 kDa protein. Substrate and inhibitor studies further support the induction of protein kinase C (PKC)-like activity. The results of N-terminal amino acid sequencing of the purified protein demonstrate identity of the kinase with an internal region of PKCδ. Immunoblot analysis was used to confirm proteolytic cleavage of intact 78 kDa PKCδ in control cells to the 40 kDa C-terminal fragment after IR exposure. The finding that both IR-induced proteolytic activation of PKCδ and endonucleolytic DNA fragmentation are blocked by Bcl-2 and Bcl-x(L) supports an association with physiological cell death (PCD). Moreover, cleavage of PKCδ occurs adjacent to aspartic acid at a site (QDN) similar to that involved in proteolytic activation of interleukin-1β converting enzyme (ICE). The specific tetrapeptide ICE inhibitor (YVAD) blocked both proteolytic activation of PKCδ and internucleosomal DNA fragmentation in IR-treated cells. These findings demonstrate that PCD is associated with proteolytic activation of PKCδ by an ICE-like protease.

Original languageEnglish (US)
Pages (from-to)6148-6156
Number of pages9
JournalEMBO Journal
Volume14
Issue number24
DOIs
StatePublished - 1995
Externally publishedYes

Keywords

  • Physiological cell death
  • Protein kinase Cδ
  • Proteolytic cleavage

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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    Emoto, Y., Manome, Y., Meinhardt, G., Kisaki, H., Kharbanda, S., Robertson, M., Ghayur, T., Wong, W. W., Kamen, R., Weichselbaum, R., & Kufe, D. (1995). Proteolytic activation of protein kinase C δ by an ICE-like protease in apoptotic cells. EMBO Journal, 14(24), 6148-6156. https://doi.org/10.1002/j.1460-2075.1995.tb00305.x