The neutrophil serine protease PR3 induces shape change of platelets via the Rho/Rho kinase and Ca2+ signaling pathways

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The neutrophil serine protease PR3 induces shape change of platelets via the Rho/Rho kinase and Ca2+ signaling pathways

Peng, Xiang (author): Linköpings universitet,Avdelningen för läkemedelsforskning,Hälsouniversitetet,Jinan University, Guangdong, China

Ramström, Sofia, 1973- (author): Linköpings universitet,Avdelningen för mikrobiologi och molekylär medicin,Hälsouniversitetet

Kurz, Tino (author): Linköpings universitet,Avdelningen för läkemedelsforskning,Hälsouniversitetet

Grenegård, Magnus, 1963- (author): Linköpings universitet,Örebro universitet,Institutionen för läkarutbildning,Department of Medical and Health Sciences, Linköping University, Linköping, Sweden;,Avdelningen för läkemedelsforskning,Hälsouniversitetet,University of Örebro, Sweden

Segelmark, Mårten (author): Östergötlands Läns Landsting,Linköpings universitet,Avdelningen för läkemedelsforskning,Hälsouniversitetet,Njurmedicinska kliniken US

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Oxon, United Kingdom : Elsevier, 2014
2014
English.
In: Thrombosis Research. - Oxon, United Kingdom : Elsevier. - 0049-3848 .- 1879-2472. ; 134:2, s. 418-425

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Journal article (peer-reviewed)

Abstract Subject headings

Introduction: Proteinase 3 (PR3) is released from neutrophil azurophilic granules and exerts complex effects on the inflammatory process. PR3 catalyzes the degradation of a number of macromolecules, but the consequences on blood cells are less well defined. In the present study, the effect of PR3 on human platelets was thoroughly investigated.Methods: The experiments were performed on washed platelets freshly isolated from blood donated by healthy human volunteers. Platelets shape change and aggregation was measured on a Chrono-Log aggregometer. The phosphorylated form of MYPT1 was visualized by immunostaining. Platelet activation was further evaluated by flow cytometry.Results: PR3 induced platelet shape change but not aggregation. Flow cytometry analysis showed that PR3 induced no P-selectin expression or binding of fibrinogen to the platelets, and it did not change the activation in response to PAR1- or PAR4-activating peptides or to thrombin. Furthermore, Fura-2 measurement and immuno-blotting analysis, respectively, revealed that PR3 stimulated small intracellular Ca2+ mobilization and Thr696-specific phosphorylation of the myosin phosphatase target subunit 1 (MYPT1). Separate treatment of platelets with the Rho/Rho kinase inhibitor Y-27632 and the intracellular Ca2+ chelator BAPTA/AM reduced the shape change induced by PR3 whereas concurrent treatment completely inhibited it.Conclusion: The data shows that the neutrophil protease PR3 is a direct modulator of human platelets and causes shape change through activation of the Rho/Rho kinase and Ca2+ signaling pathways. This finding highlights an additional mechanism in the complex interplay between neutrophils and platelets.

Subject headings

MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Hematologi (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Hematology (hsv//eng)
MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin (hsv//swe)
MEDICAL AND HEALTH SCIENCES -- Clinical Medicine (hsv//eng)

Keyword

ANCA-associated vasculitis
Proteinase PR3
Platelet shape change
Rho/Rho kinase signaling pathway
Ca2+ signaling pathway
Medicine
Medicin

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