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Glycoprotein Ib-von Willebrand Factor Interactions Activate Tyrosine Kinases in Human Platelets
Naoki Asazuma,
Yukio Ozaki,
Kaneo Satoh,
Yutaka Yatomi,
Makoto Handa,
Yoshihiro Fujimura,
Shuji Miura, and
Shoji Kume
From the Department of Clinical and Laboratory Medicine, Yamanashi Medical University, Tamaho, Nakakoma, Yamanashi, Japan; the Blood Center, Keio University, Shinjuku, Tokyo, Japan; and the Department of Blood Transfusion, Nara Medical University, Kashihara, Nara, Japan.
von Willebrand factor (vWF ) in the presence of botrocetin induces p72syk activation, assessed as its autophosphorylated level and in vitro kinase assays, the transient association of p72syk with p60c-src, and the translocation of p60c-src and p54/58lyn to cytoskeletal fractions. Jararaca glycoprotein Ib-binding protein (GPIb-BP), which specifically binds to GPIb, abolished these phenomena, suggesting that they are mediated by the vWF-GPIb interaction. These tyrosine kinase-related events were not inhibited by GRGDS peptide (plus EGTA), indicating that GPIIb/IIIa is not involved in the observed responses. Shc, an adaptor protein, was also tyrosine phosphorylated by the botrocetin-vWF activation. When GPIb was immunoprecipitated with nonfunctional monoclonal antibodies (MoAbs) directed against GPIb, a kinase activity was found to associate with GPIb upon botrocetin-vWF activation. On the other hand, anti-GPIb MoAbs that inhibit the vWF-GPIb interaction did not coprecipitate a kinase activity. Because the recovery of GPIb did not differ significantly, it is suggested that the excessive presence of inhibitory anti-GPIb MoAb dissociated a kinase activity from GPIb. Phosphoamino acid analysis showed that the kinase activity was that of a tyrosine kinase. The identity of the tyrosine kinase and the mode of interaction with the cytoplasmic region of GPIb await to be determined. Our findings suggest that the tyrosine kinase associated with GPIb serves at a most proximal step in the signal transduction pathway involved in the vWF-GPIb-induced platelet activation, which leads to other tyrosine kinase-related intracellular signals.
Blood, Vol. 90 No. 12 (December 15), 1997:
pp. 4789-4798
© 1997 by The American Society of Hematology.
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