Spontaneous neutrophil apoptosis involves caspase 3-mediated activation of protein kinase C-δ

J. Pongrácz, Paul Webb, Keqing Wang, Elizabeth Deacon, Olivia J. Lunn, Janet M. Lord

Research output: Contribution to journalArticle

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Abstract

Neutrophils are short-lived leukocytes that die by apoptosis. Whereas stress-induced apoptosis is mediated by the p38 mitogen-activated protein (MAP) kinase pathway (Frasch, S. C., Nick, J. A., Fadok, V. A., Bratton, D. L., Worthen, G. S., and Henson, P. M. (1998) J. Biol. Chem. 273, 8389-8397), signals regulating spontaneous neutrophil apoptosis have not been fully determined. In this study we found increased activation of protein kinase C (PKC)-β and -δ in neutrophils undergoing spontaneous apoptosis, but we show that only activation of PKC-δ was directly involved in the induction of apoptosis. PKC-δ can be proteolytically activated by caspase 3. We detected the 40-kDa caspase-generated fragment of PKC-δ in apoptotic neutrophils and showed that the caspase 3 inhibitor Asp-Glu-Val-Asp-fluoromethylketone prevented generation of the 40-kDa PKC-δ fragment and delayed neutrophil apoptosis. In a cell-free system, removal of PKC-δ by immunoprecipitation reduced DNA fragmentation, whereas loss of PKC-α, -β, or -ζ had no significant effect. Rottlerin and LY379196 inhibit PKC-δ and PKC-β, respectively. Only Rottlerin was able to delay neutrophil apoptosis. Inhibitors of MAP-ERK kinase 1 (PD98059) or p38 MAP kinase (SB202190) had no effect on neutrophil apoptosis, and activation of p42/44 and p38 MAP kinase did not increase in apoptotic neutrophils. We conclude that spontaneous neutrophil apoptosis involves activation of PKC-δ but is MAP kinase- independent.

Original languageEnglish
Pages (from-to)37329-37334
Number of pages6
JournalJournal of Biological Chemistry
Volume274
Issue number52
DOIs
Publication statusPublished - Dec 24 1999

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Caspase 3
Protein Kinase C
Neutrophils
Chemical activation
Apoptosis
p38 Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinases
5,21 - 12,17-dimetheneo-18H-dibenzo(i,o)pyrrolo(3,4-1)(1,8)diazacyclohexandecine-18,10(19H)dione,8((dimethylamino)methyl)-6,7,8,9,10,11-hexahydro,monomethanesulfonate
Neutrophil Activation
Caspase Inhibitors
Cell-Free System
DNA Fragmentation
Caspases
Mitogens
Immunoprecipitation
Leukocytes
Phosphotransferases
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Spontaneous neutrophil apoptosis involves caspase 3-mediated activation of protein kinase C-δ. / Pongrácz, J.; Webb, Paul; Wang, Keqing; Deacon, Elizabeth; Lunn, Olivia J.; Lord, Janet M.

In: Journal of Biological Chemistry, Vol. 274, No. 52, 24.12.1999, p. 37329-37334.

Research output: Contribution to journalArticle

Pongrácz, J. ; Webb, Paul ; Wang, Keqing ; Deacon, Elizabeth ; Lunn, Olivia J. ; Lord, Janet M. / Spontaneous neutrophil apoptosis involves caspase 3-mediated activation of protein kinase C-δ. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 52. pp. 37329-37334.
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