Proteolytic activation of protein kinase C-ε by caspase-mediated processing and transduction of antiapoptotic signals

Alakananda Basu, Dongmei Lu, Baohua Sun, Andrea N. Moor, Giridhar Rao Akkaraju, Jie Huang

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Abstract

Several novel protein kinase C (PKC) isozymes have been identified as substrates for caspase-3. We have previously shown that novel PKCε is cleaved during apoptosis in MCF-7 cells that lack any functional caspase-3. In the present study, we show that in vitro-translated PKCε is processed by human recombinant caspase-3, -7, and -9. Tumor necrosis factor-α (TNF) triggered processing of PKCε to a 43-kDa carboxyl-terminal fragment, and cell-permeable caspase inhibitors prevented TNF-induced processing of PKCε in MCF-7 cells. PKCε was cleaved primarily at the SSPD ↓ G site to generate two fragments with an approximate molecular mass of 43 kDa. It was also cleaved at the DDVD ↓ C site to generate two fragments with molecular masses of 52 and 35 kDa. Treatment of MCF-7 cells with TNF resulted in the activation of PKCε, and mutation at the SSPD ↓ G (D383A) site inhibited proteolytic activation of PKCε. Over-expression of wild-type but not dominant-negative PKCε in MCF-7 cells delayed TNF-induced apoptosis, and mutation at the D383A site prevented antiapoptotic activity of PKCε. These results suggest that cleavage of PKCε by caspase-7 at the SSPD ↓ G site results in the activation of PKCε. Furthermore, activation of PKCε was associated with its antiapoptotic function.

Original languageEnglish
Pages (from-to)41850-41856
Number of pages7
JournalJournal of Biological Chemistry
Volume277
Issue number44
DOIs
StatePublished - 1 Nov 2002

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Caspases
Protein Kinase C
Signal Transduction
Chemical activation
Processing
MCF-7 Cells
Tumor Necrosis Factor-alpha
Caspase 3
Caspase 7
Molecular mass
Apoptosis
Mutation
Caspase Inhibitors
Isoenzymes

Cite this

Basu, Alakananda ; Lu, Dongmei ; Sun, Baohua ; Moor, Andrea N. ; Akkaraju, Giridhar Rao ; Huang, Jie. / Proteolytic activation of protein kinase C-ε by caspase-mediated processing and transduction of antiapoptotic signals. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 44. pp. 41850-41856.
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Proteolytic activation of protein kinase C-ε by caspase-mediated processing and transduction of antiapoptotic signals. / Basu, Alakananda; Lu, Dongmei; Sun, Baohua; Moor, Andrea N.; Akkaraju, Giridhar Rao; Huang, Jie.

In: Journal of Biological Chemistry, Vol. 277, No. 44, 01.11.2002, p. 41850-41856.

Research output: Contribution to journalArticle

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