differential roles of the pkc novel isoforms, pkcδ and pkcε, in mouse and human platelets微分作用的pkc亚型,pkcδpkcε,老鼠和人类血小板.pdfVIP

Differential Roles of the PKC Novel Isoforms, PKCd and PKCe, in Mouse and Human Platelets 1. 1.¤a 2 2 2 Catherine J. Pears *, Kelly Thornber , Jocelyn M. Auger , Craig E. Hughes , Beata Grygielska , Majd B. Protty2, Andrew C. Pearce2¤b, Steve P. Watson2 1 Department of Biochemistry, University of Oxford, Oxford, United Kingdom, 2 Centre for Cardiovascular Sciences, Institute for Biomedical Research, Division of Medical Sciences, The Medical School, University of Birmingham, Birmingham, United Kingdom Abstract Background: Increasing evidence suggests that individual isoforms of protein kinase C (PKC) play distinct roles in regulating platelet activation. Methodology/Principal Findings: In this study, we focus on the role of two novel PKC isoforms, PKCd and PKCe, in both mouse and human platelets. PKCd is robustly expressed in human platelets and undergoes transient tyrosine phosphorylation upon stimulation by thrombin or the collagen receptor, GPVI, which becomes sustained in the presence of the pan-PKC inhibitor, Ro 31-8220. In mouse platelets, however, PKCd undergoes sustained tyrosine phosphorylation upon activation. In contrast the related isoform, PKCe, is expressed at high levels in mouse but not human platelets. There is a marked inhibition in aggregation and dense granule secretion to low concentrations of GPVI agonists in mouse platelets lacking PKCe in contrast to a minor inhibition in response to G protein-coupled receptor agonists. This reduction is mediated by inhibition of tyrosine phosphorylation of the FcRc-chain and downstream proteins, an effect also observed in wild-type mouse platelets in the presence of a PKC inhibitor