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Thromb Haemost 2015; 114(03): 558-568
DOI: 10.1160/TH14-09-0775
Cellular Haemostasis and Platelets
Schattauer GmbH

Cross talk between serine/threonine and tyrosine kinases regulates ADP-induced thromboxane generation in platelets

Dheeraj Bhavanasi
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Rachit Badolia
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Bhanu Kanth Manne
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Sumalaxmi Janapati
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Carol T. Dangelmaier
2   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Alexandra Mazharian
3   Center for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Institute of Biomedical Research, University of Birmingham, Birmingham, UK
,
Jianguo Jin
2   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Soochong Kim
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Xiaoying Zhang
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
4   Cardiovascular Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Xiongwen Chen
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
4   Cardiovascular Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Yotis A. Senis
3   Center for Cardiovascular Sciences, School of Clinical and Experimental Medicine, Institute of Biomedical Research, University of Birmingham, Birmingham, UK
,
Satya P. Kunapuli
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
5   Department 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)

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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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