CLEC-2 activates Syk through dimerization

Blood. 2010 Apr 8;115(14):2947-55. doi: 10.1182/blood-2009-08-237834. Epub 2010 Feb 12.

Abstract

The C-type lectin receptor CLEC-2 activates platelets through Src and Syk tyrosine kinases, leading to tyrosine phosphorylation of downstream adapter proteins and effector enzymes, including phospholipase-C gamma2. Signaling is initiated through phosphorylation of a single conserved tyrosine located in a YxxL sequence in the CLEC-2 cytosolic tail. The signaling pathway used by CLEC-2 shares many similarities with that used by receptors that have 1 or more copies of an immunoreceptor tyrosine-based activation motif, defined by the sequence Yxx(L/I)x(6-12)Yxx(L/I), in their cytosolic tails or associated receptor chains. Phosphorylation of the conserved immunoreceptor tyrosine-based activation motif tyrosines promotes Syk binding and activation through binding of the Syk tandem SH2 domains. In this report, we present evidence using peptide pull-down studies, surface plasmon resonance, quantitative Western blotting, tryptophan fluorescence measurements, and competition experiments that Syk activation by CLEC-2 is mediated by the cross-linking through the tandem SH2 domains with a stoichiometry of 2:1. In support of this model, cross-linking and electron microscopy demonstrate that CLEC-2 is present as a dimer in resting platelets and converted to larger complexes on activation. This is a unique mode of activation of Syk by a single YxxL-containing receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Blood Platelets / metabolism*
  • Enzyme Activation / physiology
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Lectins, C-Type / metabolism*
  • Membrane Glycoproteins / metabolism*
  • Phospholipase C gamma / metabolism
  • Platelet Activation / physiology*
  • Protein Multimerization / physiology*
  • Protein-Tyrosine Kinases / metabolism*
  • Signal Transduction / physiology*
  • Syk Kinase
  • src Homology Domains

Substances

  • CLEC2B protein, human
  • Intracellular Signaling Peptides and Proteins
  • Lectins, C-Type
  • Membrane Glycoproteins
  • Protein-Tyrosine Kinases
  • SYK protein, human
  • Syk Kinase
  • Phospholipase C gamma