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Echicetin Coated Polystyrene Beads: A Novel Tool to Investigate GPIb-Specific Platelet Activation and Aggregation [Text] / A. . Navdaev [et al.] // PLoS One. - 2014. - Vol. 9, Is. 4. - Ст. e93569, DOI 10.1371/journal.pone.0093569. - Cited References: 36. - This study was supported by DFG (SFB688, TP A2, and grant GA 1561/1-1), BMBF (01EO1003) and Rudolf-Marx-stipendium 2014 funded by Gesellschaft fur Thrombose- und Hamostaseforschung (GTH). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. . - ISSN 1932-6203

РУБ Multidisciplinary Sciences
Рубрики:
VON-WILLEBRAND-FACTOR
   GLYCOPROTEIN IB-ALPHA
   DEPENDENT PROTEIN-KINASE
   SIGNALING PATHWAY
   THROMBUS FORMATION
   INTEGRIN ALPHA(IIB)BETA(3)
   VONWILLEBRAND-FACTOR
   HIGH-SHEAR
   IGM-KAPPA
   RISTOCETIN
Аннотация: von Willebrand factor/ristocetin (vWF/R) induces GPIb-dependent platelet agglutination and activation of alpha IIb beta 3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling. Echicetin beads induced alpha IIb beta 3-dependent aggregation of washed platelets, while under the same conditions vWF/R treatment led only to alpha IIb beta 3-independent platelet agglutination. The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation, while the total amount of echicetin used for activation is not critical. Echicetin beads induced strong phosphorylation of several proteins including p38, ERK and PKB. Synergistic signaling via P2Y(12) and thromboxane receptor through secreted ADP and TxA(2), respectively, were important for echicetin bead triggered platelet activation. Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation. Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways.

WOS
Держатели документа:
[Navdaev, Alexey
Subramanian, Hariharan
Gambaryan, Stepan] Univ Wurzburg, Inst Clin Biochem & Pathobiochem, Wurzburg, Germany
[Petunin, Alexey] Russian Acad Sci, Inst Biophys, Siberian Branch, Krasnoyarsk, Russia
[Clemetson, Kenneth J.] Univ Bern, Theodor Kocher Inst, Bern, Switzerland
[Gambaryan, Stepan] Russian Acad Sci, Sechenov Inst Evolutionary Physiol & Biochem, St Petersburg 196140, Russia
[Walter, Ulrich] Univ Med Ctr Mainz, CTH, Mainz, Germany
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Navdaev, A...; Subramanian, H...; Petunin, A...; Clemetson, K.J.; Gambaryan, S...; Walter, U...; DFG [SFB688, TP A2, GA 1561/1-1]; BMBF [01EO1003]; Rudolf-Marx-stipendium; Gesellschaft fur Thrombose- und Hamostaseforschung (GTH)

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