Functional Divergence of Platelet Protein Kinase C (PKC) Isoforms in Thrombus Formation on Collagen

Karen Gilio, Matthew T. Harper, Judith M. E. M. Cosemans, Olga Konopatskaya, Imke C. A. Munnix, Lenneke Prinzen, Michael Leitges, Qinghang Liu, Jeffery D. Molkentin, Johan W. M. Heemskerk*, Alastair W. Poole

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

74 Citations (Web of Science)

Abstract

Arterial thrombosis, a major cause of myocardial infarction and stroke, is initiated by activation of blood platelets by subendothelial collagen. The protein kinase C (PKC) family centrally regulates platelet activation, and it is becoming clear that the individual PKC isoforms play distinct roles, some of which oppose each other. Here, for the first time, we address all four of the major platelet-expressed PKC isoforms, determining their comparative roles in regulating platelet adhesion to collagen and their subsequent activation under physiological flow conditions. Using mouse gene knock-out and pharmacological approaches in human platelets, we show that collagen-dependent alpha-granule secretion and thrombus formation are mediated by the conventional PKC isoforms, PKC alpha and PKC beta, whereas the novel isoform, PKC theta, negatively regulates these events. PKC delta also negatively regulates thrombus formation but not alpha-granule secretion. In addition, we demonstrate for the first time that individual PKC isoforms differentially regulate platelet calcium signaling and exposure of phosphatidylserine under flow. Although platelet deficient in PKC alpha or PKC beta showed reduced calcium signaling and phosphatidylserine exposure, these responses were enhanced in the absence of PKC theta. In summary therefore, this direct comparison between individual subtypes of PKC, by standardized methodology under flow conditions, reveals that the four major PKCs expressed in platelets play distinct non-redundant roles, where conventional PKCs promote and novel PKCs inhibit thrombus formation on collagen.
Original languageEnglish
Pages (from-to)23408-23417
JournalJournal of Biological Chemistry
Volume285
Issue number30
DOIs
Publication statusPublished - 23 Jul 2010

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