J. Exp. Med., Volume 188, Number 2, July 20, 1998 267-276

Physical and Functional Association of the Src Family Kinases Fyn and Lyn with the Collagen Receptor Glycoprotein VI-Fc Receptor  Chain Complex on Human Platelets

By Yasuharu Ezumi, Keisuke Shindoh, Masaaki Tsuji, and Hiroshi Takayama

From the Department of Hematology and Oncology, Clinical Sciences for Pathological Organs, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan

We have previously shown that uncharacterized glycoprotein VI (GPVI), which is constitutively associated and coexpressed with Fc receptor  chain (FcR) in human platelets, is essential for collagen-stimulated tyrosine phosphorylation of FcR, Syk, and phospholipase C2 (PLC2), leading to platelet activation. Here we investigated involvement of the Src family in the proximal signals through the GPVI-FcR complex, using the snake venom convulxin from Crotalus durissus terrificus, which specifically recognizes GPVI and activates platelets through cross-linking GPVI. Convulxin-coupled beads precipitated the GPVI-FcR complex from platelet lysates. Collagen and convulxin induced tyrosine phosphorylation of FcR, Syk, and PLC2 and recruited tyrosine-phosphorylated Syk to the GPVI-FcR complex. Using coprecipitation methods with convulxin-coupled beads and antibodies against FcR and the Src family, we showed that Fyn and Lyn, but not Yes, Src, Fgr, Hck, and Lck, were physically associated with the GPVI-FcR complex irrespective of stimulation. Furthermore, Fyn was rapidly activated by collagen or cross-linking GPVI. The Src family-specific inhibitor PP1 dose-dependently inhibited collagen- or convulxin-induced tyrosine phosphorylation of proteins including FcR, Syk, and PLC2, accompanied by a loss of aggregation and ATP release reaction. These results indicate that the Src family plays a critical role in platelet activation via the collagen receptor GPVI-FcR complex.

glycoprotein VI;  Fc receptor  chain;  Src family;  collagen;  platelets

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