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Thromb Haemost 2000; 83(04): 617-621
DOI: 10.1055/s-0037-1613873
Commentary
Schattauer GmbH

Identification of a Novel 33-kDa Ser/Thr Kinase that Phosphorylates the Cytoplasmic Tail of Protease-activated Receptor 1 (Thrombin Receptor) in Human Platelets

Masaru Ido
1   From the Department of Molecular Pathobiology, Mie University School of Medicine, Tsu-city, Mie, Japan
,
Tatsuya Hayashi
1   From the Department of Molecular Pathobiology, Mie University School of Medicine, Tsu-city, Mie, Japan
,
Esteban C. Gabazza
1   From the Department of Molecular Pathobiology, Mie University School of Medicine, Tsu-city, Mie, Japan
,
Koji Suzuki
1   From the Department of Molecular Pathobiology, Mie University School of Medicine, Tsu-city, Mie, Japan
› Author AffiliationsSupported in part by Grant-in-aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan (Nos. 08680680, 08557064, 09480149).
Further Information

Publication History

Received 17 January 1999

Accepted after resubmission 29 November 1999

Publication Date:
08 December 2017 (online)

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Summary

Stimulation of human platelets with thrombin or thrombin receptor agonist peptide (TRAP/ Ser-Phe-Leu-Leu-Arg-Asn) resulted in phosphorylation of the protease-activated receptor 1 (PAR1). However, protein kinase(s), capable of phosphorylating PAR1 upon activation of this receptor, has not been as yet identified in human platelets. The present study was undertaken to assess the presence of protein kinase(s) that may interact with PAR1 using a procedure based on the ability of protein kinase to undergo renaturation and phosphorylate a protein substrate fixed in a gel. We employed a fusion protein that was prepared using a glutathione S-transferase (GST) and the cytoplasmic tail of PAR1 (Pro368-Thr425)(GST-PAR1) or a reverse sequenced peptide of this domain (GST-rPAR1). The results showed that treatment of platelets with thrombin induced about 10-fold increase in the activity of the 33-kDa Ser/Thr protein kinase, which was also activated by TRAP, but not by hirudin-treated thrombin or diisopropylfluorophosphate-inactivated thrombin, suggesting that it is activated through PAR1. Furthermore, treatment of platelets with thromboxane A2 analog, STA2, led to an activation of this protein kinase and phosphorylation of PAR1. In conclusion, the present study provides evidence of homologous and heterologous activation of a novel 33-kDa Ser/Thr kinase that phosphorylates the cytoplasmic tail of PAR1.

Keywords

Platelet - protease-activated receptor 1 - thrombin - serine/threonine kinase
 

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