The Platelet Receptor CLEC-2 Is Active as a Dimer

被引：47

作者：

Watson, Aleksandra A. ^{[1
]}

Christou, Charita M. ^{[1
]}

James, John R. ^{[2
]}

Fenton-May, Angharad E. ^{[1
]}

Moncayo, Gerald E. ^{[1
]}

Mistry, Anita R. ^{[1
]}

Davis, Simon J. ^{[3
]}

Gilbert, Robert J. C. ^{[4
]}

Chakera, Aron ^{[1
]}

O'Callaghan, Chris A. ^{[1
]}

机构：

[1] Univ Oxford, Oxford OX3 7BN, England

[2] Univ Calif San Francisco, Howard Hughes Med Inst, San Francisco, CA 94158 USA

[3] Univ Oxford, John Radcliffe Hosp, Weatherall Inst Mol Med, Oxford OX3 9DS, England

[4] Div Struct Biol, Oxford OX3 7BN, England

来源：

BIOCHEMISTRY | 2009年 / 48卷 / 46期

基金：

英国医学研究理事会;

关键词：

LOW-DENSITY-LIPOPROTEIN; LIGAND-BINDING DOMAIN; CRYSTAL-STRUCTURE; LECTIN-LIKE; ALPHA(2)BETA(1) INTEGRIN; TYROSINE KINASES; ACTIVATION; PROTEIN; CELLS; GLYCOPROTEIN;

D O I：

10.1021/bi901427d

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The platelet receptor CLEC-2 binds to the snake venom toxin rhodocytin and the tumor cell surface protein podoplanin. Binding of either of these ligands promotes phosphorylation of a single tyrosine residue in the YXXL motif in the intracellular domain of CLEC-2. Phosphorylation of this tyrosine initiates binding of spleen tyrosine kinase (Syk) and triggers further downstream signaling events and ultimately potent platelet activation and aggregation. However, it is unclear how a single YXXL motif can interact efficiently with Syk, which usually recognizes two tandem YXXL repeats presented as an immunoreceptor tyrosine-based activation motif (ITAM). Using bioluminescence resonance energy transfer, coimmunopreciptitation, recombinant protein expression and analytical gel filtration chromatography, surface plasmon resonance, Western blotting, multiangle light scattering (MALS), and analytical ultracentrifugation, we show that CLEC-2 exists as a non-disulfide-linked homodimer which could allow each Syk molecule to interact with two YXXL motifs, one from each CLEC-2 monomer.

引用

页码：10988 / 10996

页数：9

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