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J. Biol. Chem., Vol. 282, Issue 5, 3165-3172, February 2, 2007

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 282/5/3165    most recent M610383200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Watson, A. A. Articles by O'Callaghan, C. A. Search for Related Content PubMed PubMed Citation Articles by Watson, A. A. Articles by O'Callaghan, C. A. Social Bookmarking                      What's this?

The Crystal Structure and Mutational Binding Analysis of the Extracellular Domain of the Platelet-activating Receptor CLEC-2^*

Aleksandra A. Watson, James Brown, Karl Harlos^¶, Johannes A. Eble^||, Thomas S. Walter^¶, and Christopher A. O'Callaghan¹

From the Henry Wellcome Building for Molecular Physiology, Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, and ^¶Oxford Protein Production Facility, Division of Structural Biology, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom and the ^||Institute for Physiological Chemistry and Pathobiochemistry, Muenster University Hospital, Waldeyerstrasse 15, 48149 Muenster, Germany

The human C-type lectin-like molecule CLEC-2 is expressed on the surface of platelets and signaling through CLEC-2 causes platelet activation and aggregation. CLEC-2 is a receptor for the platelet-aggregating snake venom protein rhodocytin. It is also a newly identified co-receptor for human immunodeficiency virus type 1 (HIV-1). An endogenous ligand has not yet been identified. We have solved the crystal structure of the extracellular domain of CLEC-2 to 1.6-Å resolution, and identified the key structural features involved in ligand binding. A semi-helical loop region and flanking residues dominate the surface that is available for ligand binding. The precise distribution of hydrophobic and electrostatic features in this loop will determine the nature of any endogenous ligand with which it can interact. Major ligand-induced conformational change in CLEC-2 is unlikely as its overall fold is compact and robust. However, ligand binding could induce a tilt of a 3–10 helical portion of the long loop region. Mutational analysis and surface plasmon resonance binding studies support these observations. This study provides a framework for understanding the effects of rhodocytin venom binding on CLEC-2 and for understanding the nature of likely endogenous ligands and will provide a basis for rational design of drugs to block ligand binding.

Received for publication, November 7, 2006

The atomic coordinates and structure factors (code 2C6U) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

^* This work was funded by the Medical Research Council. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1–S6.

¹ To whom correspondence should be addressed: Henry Wellcome Bldg. for Molecular Physiology, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, United Kingdom. Tel.: 44-1865-287789; Fax: 44-1865-287797; E-mail: chrisoc{at}ccmp.ox.ac.uk.

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