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Thromb Haemost 2013; 110(06): 1241-1249
DOI: 10.1160/TH13-03-0189
Platelets and Blood Cells
Schattauer GmbH

Trypsin causes platelet activation independently of known protease-activated receptors

Yingying Mao
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
3   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
,
Satya P. Kunapuli
1   Department of Physiology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
2   Department of Pharmacology, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
3   Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, USA
› Author Affiliations Financial support: This work is supported by the National Institutes of Health research grant HL93231 and HL118593 (S.P.K) and postdoctoral fellowship from the American Heart Association; Great Rivers affiliate (Y.M).
Further Information

Publication History

Received: 06 March 2013

Accepted after major revision: 09 August 2013

Publication Date:
30 November 2017 (online)

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Summary

To identify a physiological agonist of PAR3, we used PAR4 null murine platelets, which were known to express only PAR3. In this study, we tested several proteases and found that trypsin, but not heat-inactivated trypsin, activated PAR4 null murine platelets. Even at high concentrations, trypsin caused shape change without increasing intracellular calcium levels in PAR4 null murine platelets. Consistent with this result, the Gq inhibitor YM-254890 had no effect on trypsin-induced shape change. However, trypsin-induced platelet shape change was abolished by either p160ROCK inhibitor, Y27632 or H1152. Furthermore, trypsin caused phosphorylation of myosin light chain (Thr18), but not Akt or Erk. Surprisingly, trypsin caused a similar shape change in PAR4-desensitised PAR3 null murine platelets as in PAR4 null murine platelets, indicating that trypsin did not activate PAR3 to cause shape change. More interestingly, the Src family kinase (SFK) inhibitor PP2 abolished trypsin-induced, but not AYPGKF-induced, shape change. Hence, trypsin activated a novel signalling pathway through RhoA/p160ROCK and was regulated by SFKs. In conclusion, our study demonstrates a novel protease signalling pathway in platelets that is independent of PARs. This protease-induced novel signalling pathway regulates platelet shape change through SFKs and p160ROCK.

Keywords

Trypsin - protease-activated receptor - platelet shape change
 

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