Identification and characterization of protein phosphatase 2C activation by ceramide

David M. Perry, Kazuyuki Kitatani, Patrick Roddy, Mohamad El-Osta, Yusuf A. Hannun

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

Ceramide is a bioactive sphingolipid with many associated biological outcomes, yet there is a significant gap in our current understanding of how ceramide mediates these processes. Previously, ceramide has been shown to activate protein phosphatase (PP) 1 and 2A. While continuing this line of work, a late fraction from a Mono-Q column was consistently observed to be activated by ceramide, yet PP1 and PP2A were undetectable in this fraction. Proteomic analysis of this fraction revealed the identity of the phosphatase to be PP2Cγ/PPM1G. This was consistent with our findings that PP2Cγ 1-eluted in a high salt fraction due to its strongly acidic domain, and 2-was insensitive to okadaic acid. Further characterization was performed with PP2Cα, which showed robust activation by C6-ceramide. Activation was specific for the erythro conformation of ceramide and the presence of the acyl chain and hydroxyl group at the first carbon. In order to demonstrate more physiological activation of PP2Cα by ceramide, phospho-p38δ was utilized as substrate. Indeed, PP2Cα induced the dephosphorylation of p38δ only in the presence of C16-ceramide. Taken together, these results show that the PP2C family of phosphatases is activated by ceramide, which may have important consequences in mediating the biological effects of ceramide.

Original languageEnglish
Pages (from-to)1513-1521
Number of pages9
JournalJournal of lipid research
Volume53
Issue number8
DOIs
Publication statusPublished - 2012 Aug

Keywords

  • PP2C
  • Ser/Thr phosphatases
  • Sphingolipids

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Fingerprint Dive into the research topics of 'Identification and characterization of protein phosphatase 2C activation by ceramide'. Together they form a unique fingerprint.

Cite this