A Novel Viper Venom Metalloproteinase, Alborhagin, Is an Agonist at the Platelet Collagen Receptor GPVI

Robert K. Andrews, Elizabeth E. Gardiner, Naoki Asazuma, Oscar Berlanga, David Tulasne, Bernhard Nieswandtll, A. Ian Smith, Michael C. Berndt, Stephen P. Watson

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Abstract

The interaction of platelet membrane glycoprotein VI (GPVI) with collagen can initiate (patho)physiological thrombus formation. The viper venom C-type lectin family proteins convulxin and alboaggregin-A activate platelets by interacting with GPVI. In this study, we isolated from white-lipped tree viper (Trimeresurus albolabris) venom, alborhagin, which is functionally related to convulxin because it activates platelets but is structurally different and related to venom metallopro. teinases. Alborhagin-induced platelet aggregation (EC50, <7.5 μg/ml) was inhibitable by an anti-αIIbβ 3 antibody, CRC64, and the Src family kinase inhibitor PP1, suggesting that alborhagin activates platelets, leading to αIIbβ3-dependent aggregation. Additional evidence suggested that, like convulxin, alborhagin activated platelets by a mechanism involving GPVI. First, alborhagin- and convulxin-treated platelets showed a similar tyrosine phosphorylation pattern, including a similar level of phospholipase Cγ2 phosphorylation. Second, alborhagin induced GPVI-dependent responses in GPVI-transfected K562 and Jurkat cells. Third, alborhagin-dependent aggregation of mouse platelets was inhibited by the anti-GPVI monoclonal antibody JAQ1. Alborhagin had minimal effect on convulxin binding to GPVI-expressing cells, indicating that these venom proteins may recognize distinct binding sites. Characterization of alborhagin as a GPVI agonist that is structurally distinct from convulxin demonstrates the versatility of snake venom toxins and provides a novel probe for GPVI-dependent platelet activation.

Original languageEnglish
Pages (from-to)28092-28097
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number30
DOIs
Publication statusPublished - 27 Jul 2001
Externally publishedYes

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