Dissociation of protein kinase C activation and sn-1,2-diacylglycerol formation: Comparison of phosphatidylinositol- and phosphatidylcholine-derived diglycerides in α-thrombin-stimulated fibroblasts

K. L. Leach, V. A. Ruff, T. M. Wright, M. S. Pessin, D. M. Raben

Research output: Contribution to journalArticle

Abstract

Diacylglycerols (DAGs) derived from phosphatidylcholine (PC) hydrolysis have been shown to activate protein kinase C (PKC) in vitro, but it is not known whether this event occurs in response to DAGs generated via agonist-induced PC hydrolysis in intact cells. In this report we have addressed this question directly, using α-thrombin stimulation of IIC9 fibroblasts. PKC activation in intact cells was assessed in two ways, by measuring: 1) PKC membrane association as determined by kinase activity and Western blot analysis and 2) the phosphorylation of an endogenous PKC substrate, an 80-kDa protein. Treatment with 500 ng/ml α-thrombin has been shown to stimulate both phosphoinositide and PC hydrolysis, whereas treatment with 100 pg/ml α-thrombin stimulates only PC breakdown. Using these two conditions, we show that DAG produced from phosphoinositide, but not PC hydrolysis, is associated with the activation of PKC.

Original languageEnglish (US)
Pages (from-to)3215-3221
Number of pages7
JournalJournal of Biological Chemistry
Volume266
Issue number5
StatePublished - Jul 10 1991

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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