Cross talk between serine/threonine and tyrosine kinases regulates ADP-induced thromboxane generation in platelets (original) (raw)
Thromb Haemost 2015; 114(03): 558-568
DOI: 10.1160/TH14-09-0775
Cellular Haemostasis and Platelets
Schattauer GmbH
Dheeraj Bhavanasi
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
Rachit Badolia
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Bhanu Kanth Manne
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Sumalaxmi Janapati
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Carol T. Dangelmaier
2Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Alexandra Mazharian
3Center for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Institute of Biomedical Research, University of Birmingham, Birmingham, UK
,
Jianguo Jin
2Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Soochong Kim
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Xiaoying Zhang
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
4Cardiovascular Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Xiongwen Chen
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
4Cardiovascular Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Yotis A. Senis
3Center for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Institute of Biomedical Research, University of Birmingham, Birmingham, UK
,
Satya P. Kunapuli
1Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
5Department of Pharmacology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
› Author Affiliations Financial support: This work is supported by HL93231 and HL118593 from National Institutes of Health to SPK.
Further Information
Publication History
Received: 16 September 2014
Accepted after major revision: 26 March 2015
Publication Date:
21 November 2017 (online)
Summary
ADP-induced thromboxane generation depends on Src family kinases (SFKs) and is enhanced with pan-protein kinase C (PKC) inhibitors, but it is not clear how these two events are linked. The aim of the current study is to investigate the role of Y311 phosphorylated PKCδ in regulating ADP-induced platelet activation. In the current study, we employed various inhibitors and murine platelets from mice deficient in specific molecules to evaluate the role of PKCδ in ADP-induced platelet responses. We show that, upon stimulation of platelets with 2MeSADP, Y311 on PKCδ is phosphorylated in a P2Y1/Gq and Lyn-dependent manner. By using PKCδ and Lyn knockout murine platelets, we also show that tyrosine phosphorylated PKCδ plays a functional role in mediating 2MeSADP-induced thromboxane generation. 2MeSADP-induced PKCδ Y311 phosphorylation and thromboxane generation were potentiated in human platelets pre-treated with either a pan-PKC inhibitor, GF109203X or a PKC α/β inhibitor and in PKC α or β knockout murine platelets compared to controls. Furthermore, we show that PKC α/β inhibition potentiates the activity of SFK, which further hyper-phosphorylates PKCδ and potentiates thromboxane generation. These results show for the first time that tyrosine phosphorylated PKCδ regulates ADP-induced thromboxane generation independent of its catalytic activity and that classical PKC isoforms α/β regulate the tyrosine phosphorylation on PKCδ and subsequent thromboxane generation through tyrosine kinase, Lyn, in platelets.
Keywords
Platelet physiology - protein kinase C - thromboxane A2 - ADP receptors - protein tyrosine kinases
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