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

Dheeraj Bhavanasi; Rachit Badolia; Bhanu Kanth Manne; Sumalaxmi Janapati; Carol T. Dangelmaier; Alexandra Mazharian; Jianguo Jin; Soochong Kim; Xiaoying Zhang; Xiongwen Chen; Yotis A. Senis; Satya P. Kunapuli

doi:10.1160/TH14-09-0775

Thrombosis and Haemostasis, Table of Contents

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

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

Abstract

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

Full Text

References

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