GPVI potentiation of platelet activation by thrombin and adhesion molecules independent of Src kinases and Syk

Sascha C. Hughan, Craig E. Hughes, Owen McCarty, Edina Schweighoffer, Izoumroud Soultanova, Jerry Ware, Victor L J Tybulewicz, Steve P. Watson

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

OBJECTIVE - The present study investigates the role of Src and Syk tyrosine kinases in signaling by G-protein coupled and platelet adhesion receptors. METHODS AND RESULTS - Using Syk platelets or the Src kinase inhibitor PP2, we demonstrate a critical role for Src and Syk kinases in mediating lamellipodia formation on VWF, collagen, CRP, fibrinogen, and fibronectin. In all cases, the spreading defect was overcome by addition of thrombin. Conversely, platelet aggregation and αIIbβ3 activation induced by thrombin was similar to controls, arguing against a functional role for Src and Syk in αIIbβ3 activation. Unexpectedly, CRP potentiated integrin αIIbβ3 activation and platelet aggregation induced by subthreshold concentrations of thrombin in Syk platelets or in the presence of the Src kinase inhibitor PP2. Potentiation in the presence of PP2 was lost in the absence of FcRγ-chain or GPVI confirming that it was mediated through the immunoglobulin receptor. Further delineation of this PP2-resistant synergy revealed that PAR4 could trigger the enhanced response in combination with CRP. CONCLUSIONS - We show that Syk is critical for lamellipodia formation on a range of immobilized proteins but that this can be overcome by addition of thrombin. Further, we reveal a novel role for GPVI in supporting thrombin-induced activation, independent of Syk and Src kinases.

Original languageEnglish (US)
Pages (from-to)422-429
Number of pages8
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume27
Issue number2
DOIs
StatePublished - Feb 2007

Fingerprint

src-Family Kinases
Platelet Activation
Thrombin
Pseudopodia
Blood Platelets
Platelet Aggregation
Immobilized Proteins
Fibronectins
GTP-Binding Proteins
Integrins
Fibrinogen
Immunoglobulins
Collagen
Syk Kinase

Keywords

  • GPVI signaling
  • Platelets
  • Src kinases
  • Syk
  • Thrombin

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

GPVI potentiation of platelet activation by thrombin and adhesion molecules independent of Src kinases and Syk. / Hughan, Sascha C.; Hughes, Craig E.; McCarty, Owen; Schweighoffer, Edina; Soultanova, Izoumroud; Ware, Jerry; Tybulewicz, Victor L J; Watson, Steve P.

In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 27, No. 2, 02.2007, p. 422-429.

Research output: Contribution to journalArticle

Hughan, Sascha C. ; Hughes, Craig E. ; McCarty, Owen ; Schweighoffer, Edina ; Soultanova, Izoumroud ; Ware, Jerry ; Tybulewicz, Victor L J ; Watson, Steve P. / GPVI potentiation of platelet activation by thrombin and adhesion molecules independent of Src kinases and Syk. In: Arteriosclerosis, Thrombosis, and Vascular Biology. 2007 ; Vol. 27, No. 2. pp. 422-429.
@article{1801db03ece4422ebfe24d1bff061729,
title = "GPVI potentiation of platelet activation by thrombin and adhesion molecules independent of Src kinases and Syk",
abstract = "OBJECTIVE - The present study investigates the role of Src and Syk tyrosine kinases in signaling by G-protein coupled and platelet adhesion receptors. METHODS AND RESULTS - Using Syk platelets or the Src kinase inhibitor PP2, we demonstrate a critical role for Src and Syk kinases in mediating lamellipodia formation on VWF, collagen, CRP, fibrinogen, and fibronectin. In all cases, the spreading defect was overcome by addition of thrombin. Conversely, platelet aggregation and αIIbβ3 activation induced by thrombin was similar to controls, arguing against a functional role for Src and Syk in αIIbβ3 activation. Unexpectedly, CRP potentiated integrin αIIbβ3 activation and platelet aggregation induced by subthreshold concentrations of thrombin in Syk platelets or in the presence of the Src kinase inhibitor PP2. Potentiation in the presence of PP2 was lost in the absence of FcRγ-chain or GPVI confirming that it was mediated through the immunoglobulin receptor. Further delineation of this PP2-resistant synergy revealed that PAR4 could trigger the enhanced response in combination with CRP. CONCLUSIONS - We show that Syk is critical for lamellipodia formation on a range of immobilized proteins but that this can be overcome by addition of thrombin. Further, we reveal a novel role for GPVI in supporting thrombin-induced activation, independent of Syk and Src kinases.",
keywords = "GPVI signaling, Platelets, Src kinases, Syk, Thrombin",
author = "Hughan, {Sascha C.} and Hughes, {Craig E.} and Owen McCarty and Edina Schweighoffer and Izoumroud Soultanova and Jerry Ware and Tybulewicz, {Victor L J} and Watson, {Steve P.}",
year = "2007",
month = "2",
doi = "10.1161/01.ATV.0000252826.96134.21",
language = "English (US)",
volume = "27",
pages = "422--429",
journal = "Arteriosclerosis, Thrombosis, and Vascular Biology",
issn = "1079-5642",
publisher = "Lippincott Williams and Wilkins",
number = "2",

}

TY - JOUR

T1 - GPVI potentiation of platelet activation by thrombin and adhesion molecules independent of Src kinases and Syk

AU - Hughan, Sascha C.

AU - Hughes, Craig E.

AU - McCarty, Owen

AU - Schweighoffer, Edina

AU - Soultanova, Izoumroud

AU - Ware, Jerry

AU - Tybulewicz, Victor L J

AU - Watson, Steve P.

PY - 2007/2

Y1 - 2007/2

N2 - OBJECTIVE - The present study investigates the role of Src and Syk tyrosine kinases in signaling by G-protein coupled and platelet adhesion receptors. METHODS AND RESULTS - Using Syk platelets or the Src kinase inhibitor PP2, we demonstrate a critical role for Src and Syk kinases in mediating lamellipodia formation on VWF, collagen, CRP, fibrinogen, and fibronectin. In all cases, the spreading defect was overcome by addition of thrombin. Conversely, platelet aggregation and αIIbβ3 activation induced by thrombin was similar to controls, arguing against a functional role for Src and Syk in αIIbβ3 activation. Unexpectedly, CRP potentiated integrin αIIbβ3 activation and platelet aggregation induced by subthreshold concentrations of thrombin in Syk platelets or in the presence of the Src kinase inhibitor PP2. Potentiation in the presence of PP2 was lost in the absence of FcRγ-chain or GPVI confirming that it was mediated through the immunoglobulin receptor. Further delineation of this PP2-resistant synergy revealed that PAR4 could trigger the enhanced response in combination with CRP. CONCLUSIONS - We show that Syk is critical for lamellipodia formation on a range of immobilized proteins but that this can be overcome by addition of thrombin. Further, we reveal a novel role for GPVI in supporting thrombin-induced activation, independent of Syk and Src kinases.

AB - OBJECTIVE - The present study investigates the role of Src and Syk tyrosine kinases in signaling by G-protein coupled and platelet adhesion receptors. METHODS AND RESULTS - Using Syk platelets or the Src kinase inhibitor PP2, we demonstrate a critical role for Src and Syk kinases in mediating lamellipodia formation on VWF, collagen, CRP, fibrinogen, and fibronectin. In all cases, the spreading defect was overcome by addition of thrombin. Conversely, platelet aggregation and αIIbβ3 activation induced by thrombin was similar to controls, arguing against a functional role for Src and Syk in αIIbβ3 activation. Unexpectedly, CRP potentiated integrin αIIbβ3 activation and platelet aggregation induced by subthreshold concentrations of thrombin in Syk platelets or in the presence of the Src kinase inhibitor PP2. Potentiation in the presence of PP2 was lost in the absence of FcRγ-chain or GPVI confirming that it was mediated through the immunoglobulin receptor. Further delineation of this PP2-resistant synergy revealed that PAR4 could trigger the enhanced response in combination with CRP. CONCLUSIONS - We show that Syk is critical for lamellipodia formation on a range of immobilized proteins but that this can be overcome by addition of thrombin. Further, we reveal a novel role for GPVI in supporting thrombin-induced activation, independent of Syk and Src kinases.

KW - GPVI signaling

KW - Platelets

KW - Src kinases

KW - Syk

KW - Thrombin

UR - http://www.scopus.com/inward/record.url?scp=33846440543&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33846440543&partnerID=8YFLogxK

U2 - 10.1161/01.ATV.0000252826.96134.21

DO - 10.1161/01.ATV.0000252826.96134.21

M3 - Article

C2 - 17110603

AN - SCOPUS:33846440543

VL - 27

SP - 422

EP - 429

JO - Arteriosclerosis, Thrombosis, and Vascular Biology

JF - Arteriosclerosis, Thrombosis, and Vascular Biology

SN - 1079-5642

IS - 2

ER -