The platelet receptor CLEC-2 is active as a dimer

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 ini...

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Main Authors: Watson, A., Christou, C., James, J., Fenton-May, A., Moncayo, G., Mistry, A., Davis, S., Gilbert, R., Chakera, Aron, O'Callaghan, C.
Format: Journal Article
Published: American Chemical Society 2009
Online Access:http://hdl.handle.net/20.500.11937/4827
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author Watson, A.
Christou, C.
James, J.
Fenton-May, A.
Moncayo, G.
Mistry, A.
Davis, S.
Gilbert, R.
Chakera, Aron
O'Callaghan, C.
author_facet Watson, A.
Christou, C.
James, J.
Fenton-May, A.
Moncayo, G.
Mistry, A.
Davis, S.
Gilbert, R.
Chakera, Aron
O'Callaghan, C.
author_sort Watson, A.
building Curtin Institutional Repository
collection Online Access
description 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, coimmuno-preciptitation, 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 alloweach Syk molecule to interact with two YXXL motifs, one from each CLEC-2 monomer. © 2009 American Chemical Society.
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spelling curtin-20.500.11937-48272017-09-13T14:46:23Z The platelet receptor CLEC-2 is active as a dimer Watson, A. Christou, C. James, J. Fenton-May, A. Moncayo, G. Mistry, A. Davis, S. Gilbert, R. Chakera, Aron O'Callaghan, C. 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, coimmuno-preciptitation, 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 alloweach Syk molecule to interact with two YXXL motifs, one from each CLEC-2 monomer. © 2009 American Chemical Society. 2009 Journal Article http://hdl.handle.net/20.500.11937/4827 10.1021/bi901427d American Chemical Society restricted
spellingShingle Watson, A.
Christou, C.
James, J.
Fenton-May, A.
Moncayo, G.
Mistry, A.
Davis, S.
Gilbert, R.
Chakera, Aron
O'Callaghan, C.
The platelet receptor CLEC-2 is active as a dimer
title The platelet receptor CLEC-2 is active as a dimer
title_full The platelet receptor CLEC-2 is active as a dimer
title_fullStr The platelet receptor CLEC-2 is active as a dimer
title_full_unstemmed The platelet receptor CLEC-2 is active as a dimer
title_short The platelet receptor CLEC-2 is active as a dimer
title_sort platelet receptor clec-2 is active as a dimer
url http://hdl.handle.net/20.500.11937/4827