Phosphoinositide interconversion in thrombin-stimulated human platelets

D. B. Wilson, E. J. Neufeld, P. W. Majerus

Research output: Contribution to journalArticle

116 Scopus citations

Abstract

Stimulation of platelets and other secretory cells by agonists results in the degradation of phosphoinositides by phospholipase C. Kinetic studies suggest that hydrolysis of phosphatidylinositol 4,5-diphosphate (PI-4,5-P2) is an initial event in this process. Platelets contain much larger amounts of phosphatidylinositol (PI) than PI-4,5-P2, and approximately 50% of total phosphoinositides are degraded upon stimulation. We have investigated whether degradation of PI occurs by direct phospholipase C hydrolysis or by phosphorylation to PI-4,5-P2 followed by phospholipase C action on the latter compound. When platelets are incubated for 3 min with 32P(i) prior to stimulation, the phosphoinositides are labeled to different specific activities. Under these nonequilibrium conditions, the time course of change in specific activity reflects turnover. The rise in specific activity of phosphatidylinositol 4-phosphate (PI-4-P) is similar in stimulated and unstimulated cells, indicating that there is little increase in the conversion of PI to PI-4-P during thrombin stimulation. In addition, the specific activity of the 4-phosphate in PI-4-P during thrombin stimulation is less than both the 5-phosphate of PI-4,5-P2 and the phosphate group of phosphatidic acid, indicating that the 4-phosphate moiety is not labeled to equilibrium with ATP. This finding is inconsistent with a rapid flux of PI via PI-4-P to PI-4,5-P2 during thrombin stimulation, in which case the 4-phosphate would be at maximum specific activity. We, therefore, conclude that the bulk of PI breakdown that occurs in thrombin-stimulated platelets occurs via direct phospholipase C hydrolysis of PI.

Original languageEnglish
Pages (from-to)1046-1051
Number of pages6
JournalJournal of Biological Chemistry
Volume260
Issue number2
StatePublished - Jan 1 1985

Fingerprint Dive into the research topics of 'Phosphoinositide interconversion in thrombin-stimulated human platelets'. Together they form a unique fingerprint.

  • Cite this