A role for glycosphingolipid-enriched microdomains in platelet glycoprotein Ib-mediated platelet activation

W. Jin, O. Inoue, N. Tamura, K. Suzuki-inoue, K. Satoh, M. C. Berndt, M. Handa, S. Goto, Yukio Ozaki

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Background: Glycoprotein (GP) Ib, a platelet von Willebrand factor (VWF) receptor, plays a crucial role in thrombosis and hemostasis. As recent reports have suggested that GPIb partially locates in a particular region, designated as glycosphingolipid-enriched microdomains (GEMs), we hypothesized that GEMs play a central role in GPIb-mediated platelet activation. Methods: Platelets were stimulated by VWF/botrocetin to activate platelets through GPIb. GEMs and non-GEMs were isolated by sucrose density gradient ultracentrifugation and the location of signaling molecules characterized. The role of GEMs-mediated signaling in platelet behavior was tested by platelet aggregation and by platelet interaction with immobilized VWF under flow conditions when GEMs were disrupted by methyl-β-cyclodextrin (MβCD). Results: GPIb was partially translocated to GEMs upon VWF/botrocetin stimulation. Immunoprecipitation of GPIb in GEMs and non-GEMs revealed that the tyrosine kinases, Src and Lyn, were associated with GPIb only in GEMs after GPIb-stimulation, and not in non-GEMs. Activation of PLCγ2 was more intense in GEMs than non-GEMs. Disruption of GEMs by MβCD strongly inhibited tyrosine phosphorylation of Syk and PLCγ2. Functional studies revealed that stable adhesion of platelets to a VWF-coated surface under flow was impaired by GEM disruption by MβCD. Conclusion: The combined results suggest that GEMs play an important role in GPIb-mediated platelet activation.

Original languageEnglish
Pages (from-to)1034-1040
Number of pages7
JournalJournal of Thrombosis and Haemostasis
Volume5
Issue number5
DOIs
Publication statusPublished - 2007 May

Fingerprint

Platelet Glycoprotein GPIb-IX Complex
Platelet Membrane Glycoproteins
Glycosphingolipids
Platelet Activation
Blood Platelets
von Willebrand Factor
Cyclodextrins
src-Family Kinases
Ultracentrifugation
Hemostasis
Platelet Aggregation
Immunoprecipitation
Tyrosine
Sucrose

Keywords

  • GEMs
  • GPIb
  • Lyn
  • Src

ASJC Scopus subject areas

  • Medicine(all)

Cite this

A role for glycosphingolipid-enriched microdomains in platelet glycoprotein Ib-mediated platelet activation. / Jin, W.; Inoue, O.; Tamura, N.; Suzuki-inoue, K.; Satoh, K.; Berndt, M. C.; Handa, M.; Goto, S.; Ozaki, Yukio.

In: Journal of Thrombosis and Haemostasis, Vol. 5, No. 5, 05.2007, p. 1034-1040.

Research output: Contribution to journalArticle

Jin, W, Inoue, O, Tamura, N, Suzuki-inoue, K, Satoh, K, Berndt, MC, Handa, M, Goto, S & Ozaki, Y 2007, 'A role for glycosphingolipid-enriched microdomains in platelet glycoprotein Ib-mediated platelet activation', Journal of Thrombosis and Haemostasis, vol. 5, no. 5, pp. 1034-1040. https://doi.org/10.1111/j.1538-7836.2007.02476.x
Jin, W. ; Inoue, O. ; Tamura, N. ; Suzuki-inoue, K. ; Satoh, K. ; Berndt, M. C. ; Handa, M. ; Goto, S. ; Ozaki, Yukio. / A role for glycosphingolipid-enriched microdomains in platelet glycoprotein Ib-mediated platelet activation. In: Journal of Thrombosis and Haemostasis. 2007 ; Vol. 5, No. 5. pp. 1034-1040.
@article{ecf09caee8484fb1ab3f3c4299a166e3,
title = "A role for glycosphingolipid-enriched microdomains in platelet glycoprotein Ib-mediated platelet activation",
abstract = "Background: Glycoprotein (GP) Ib, a platelet von Willebrand factor (VWF) receptor, plays a crucial role in thrombosis and hemostasis. As recent reports have suggested that GPIb partially locates in a particular region, designated as glycosphingolipid-enriched microdomains (GEMs), we hypothesized that GEMs play a central role in GPIb-mediated platelet activation. Methods: Platelets were stimulated by VWF/botrocetin to activate platelets through GPIb. GEMs and non-GEMs were isolated by sucrose density gradient ultracentrifugation and the location of signaling molecules characterized. The role of GEMs-mediated signaling in platelet behavior was tested by platelet aggregation and by platelet interaction with immobilized VWF under flow conditions when GEMs were disrupted by methyl-β-cyclodextrin (MβCD). Results: GPIb was partially translocated to GEMs upon VWF/botrocetin stimulation. Immunoprecipitation of GPIb in GEMs and non-GEMs revealed that the tyrosine kinases, Src and Lyn, were associated with GPIb only in GEMs after GPIb-stimulation, and not in non-GEMs. Activation of PLCγ2 was more intense in GEMs than non-GEMs. Disruption of GEMs by MβCD strongly inhibited tyrosine phosphorylation of Syk and PLCγ2. Functional studies revealed that stable adhesion of platelets to a VWF-coated surface under flow was impaired by GEM disruption by MβCD. Conclusion: The combined results suggest that GEMs play an important role in GPIb-mediated platelet activation.",
keywords = "GEMs, GPIb, Lyn, Src",
author = "W. Jin and O. Inoue and N. Tamura and K. Suzuki-inoue and K. Satoh and Berndt, {M. C.} and M. Handa and S. Goto and Yukio Ozaki",
year = "2007",
month = "5",
doi = "10.1111/j.1538-7836.2007.02476.x",
language = "English",
volume = "5",
pages = "1034--1040",
journal = "Journal of Thrombosis and Haemostasis",
issn = "1538-7933",
publisher = "Wiley-Blackwell",
number = "5",

}

TY - JOUR

T1 - A role for glycosphingolipid-enriched microdomains in platelet glycoprotein Ib-mediated platelet activation

AU - Jin, W.

AU - Inoue, O.

AU - Tamura, N.

AU - Suzuki-inoue, K.

AU - Satoh, K.

AU - Berndt, M. C.

AU - Handa, M.

AU - Goto, S.

AU - Ozaki, Yukio

PY - 2007/5

Y1 - 2007/5

N2 - Background: Glycoprotein (GP) Ib, a platelet von Willebrand factor (VWF) receptor, plays a crucial role in thrombosis and hemostasis. As recent reports have suggested that GPIb partially locates in a particular region, designated as glycosphingolipid-enriched microdomains (GEMs), we hypothesized that GEMs play a central role in GPIb-mediated platelet activation. Methods: Platelets were stimulated by VWF/botrocetin to activate platelets through GPIb. GEMs and non-GEMs were isolated by sucrose density gradient ultracentrifugation and the location of signaling molecules characterized. The role of GEMs-mediated signaling in platelet behavior was tested by platelet aggregation and by platelet interaction with immobilized VWF under flow conditions when GEMs were disrupted by methyl-β-cyclodextrin (MβCD). Results: GPIb was partially translocated to GEMs upon VWF/botrocetin stimulation. Immunoprecipitation of GPIb in GEMs and non-GEMs revealed that the tyrosine kinases, Src and Lyn, were associated with GPIb only in GEMs after GPIb-stimulation, and not in non-GEMs. Activation of PLCγ2 was more intense in GEMs than non-GEMs. Disruption of GEMs by MβCD strongly inhibited tyrosine phosphorylation of Syk and PLCγ2. Functional studies revealed that stable adhesion of platelets to a VWF-coated surface under flow was impaired by GEM disruption by MβCD. Conclusion: The combined results suggest that GEMs play an important role in GPIb-mediated platelet activation.

AB - Background: Glycoprotein (GP) Ib, a platelet von Willebrand factor (VWF) receptor, plays a crucial role in thrombosis and hemostasis. As recent reports have suggested that GPIb partially locates in a particular region, designated as glycosphingolipid-enriched microdomains (GEMs), we hypothesized that GEMs play a central role in GPIb-mediated platelet activation. Methods: Platelets were stimulated by VWF/botrocetin to activate platelets through GPIb. GEMs and non-GEMs were isolated by sucrose density gradient ultracentrifugation and the location of signaling molecules characterized. The role of GEMs-mediated signaling in platelet behavior was tested by platelet aggregation and by platelet interaction with immobilized VWF under flow conditions when GEMs were disrupted by methyl-β-cyclodextrin (MβCD). Results: GPIb was partially translocated to GEMs upon VWF/botrocetin stimulation. Immunoprecipitation of GPIb in GEMs and non-GEMs revealed that the tyrosine kinases, Src and Lyn, were associated with GPIb only in GEMs after GPIb-stimulation, and not in non-GEMs. Activation of PLCγ2 was more intense in GEMs than non-GEMs. Disruption of GEMs by MβCD strongly inhibited tyrosine phosphorylation of Syk and PLCγ2. Functional studies revealed that stable adhesion of platelets to a VWF-coated surface under flow was impaired by GEM disruption by MβCD. Conclusion: The combined results suggest that GEMs play an important role in GPIb-mediated platelet activation.

KW - GEMs

KW - GPIb

KW - Lyn

KW - Src

UR - http://www.scopus.com/inward/record.url?scp=34247892465&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34247892465&partnerID=8YFLogxK

U2 - 10.1111/j.1538-7836.2007.02476.x

DO - 10.1111/j.1538-7836.2007.02476.x

M3 - Article

VL - 5

SP - 1034

EP - 1040

JO - Journal of Thrombosis and Haemostasis

JF - Journal of Thrombosis and Haemostasis

SN - 1538-7933

IS - 5

ER -