Direct binding to ceramide activates protein kinase Cζ before the formation of a pro-apoptotic complex with PAR-4 in differentiating stem cells

Guanghu Wang, Maria J Silva, Kannan Krishnamurthy, Eric Tran, Brian G. Condie, Erhard Bieberich

Research output: Contribution to journalArticle

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Abstract

We have reported that ceramide mediates binding of atypical protein kinase C (PKC) ζ to its inhibitor protein, PAR-4 (prostate apoptosis response-4), thereby inducing apoptosis in differentiating embryonic stem cells. Using a novel method of lipid vesicle-mediated affinity chromatography, we showed here that endogenous ceramide binds directly to the PKCζ-PAR-4 complex. Ceramide and its analogs activated PKCζ prior to binding to PAR-4, as determined by increased levels of phosphorylated PKCζ and glycogen synthase kinase-3β and emergence of a PAR-4-to-phosphorylated PKCζ fluorescence resonance energy transfer signal that co-localizes with ceramide. Elevated expression and activation of PKCζ increased cell survival, whereas expression of PAR-4 promoted apoptosis. This suggests that PKCζ counteracts apoptosis, unless its ceramide-induced activation is compromised by binding to PAR-4. A luciferase reporter assay showed that ceramide analogs activate nuclear factor (NF)-κB unless PAR-4-dependent inhibition of PKCζ suppresses NF-κB activation. Taken together, our results show that direct physical association with ceramide and PAR-4 regulates the activity of PKCζ. They also indicate that this interaction regulates the activity of glycogen synthase kinase-3β and NF-κB.

Original languageEnglish (US)
Pages (from-to)26415-26424
Number of pages10
JournalJournal of Biological Chemistry
Volume280
Issue number28
DOIs
StatePublished - Jul 15 2005

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Ceramides
Stem cells
Protein Kinase C
Prostate
Stem Cells
Apoptosis
Glycogen Synthase Kinase 3
Chemical activation
Inhibitor of Apoptosis Proteins
Affinity chromatography
Fluorescence Resonance Energy Transfer
Embryonic Stem Cells
Luciferases
Affinity Chromatography
Assays
Cell Survival
Carrier Proteins
Cells
Association reactions
Lipids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Direct binding to ceramide activates protein kinase Cζ before the formation of a pro-apoptotic complex with PAR-4 in differentiating stem cells. / Wang, Guanghu; Silva, Maria J; Krishnamurthy, Kannan; Tran, Eric; Condie, Brian G.; Bieberich, Erhard.

In: Journal of Biological Chemistry, Vol. 280, No. 28, 15.07.2005, p. 26415-26424.

Research output: Contribution to journalArticle

Wang, Guanghu ; Silva, Maria J ; Krishnamurthy, Kannan ; Tran, Eric ; Condie, Brian G. ; Bieberich, Erhard. / Direct binding to ceramide activates protein kinase Cζ before the formation of a pro-apoptotic complex with PAR-4 in differentiating stem cells. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 28. pp. 26415-26424.
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