Molecular priming of Lyn by GPVI enables an immune receptor to adopt a hemostatic role

The immune receptor signaling pathway is used by nonimmune cells, but the molecular adaptations that underlie its functional diversification are not known. Circulating platelets use the immune receptor homologue glycoprotein VI (GPVI) to respond to collagen exposed at sites of vessel injury. In contrast to immune cell responses, platelet activation must take place within seconds to successfully form thrombi in flowing blood. Here, we show that the GPVI receptor utilizes a unique intracellular proline-rich domain (PRD) to accelerate platelet activation, a requirement for efficient platelet adhesion to collagen under flow. The GPVI PRD specifically binds the Src-family kinase Lyn and directly activates it, presumably through SH3 displacement. In resting platelets, Lyn is constitutively bound to GPVI in an activated state and platelets lacking Lyn exhibit defective collagen adhesion like that of platelets with GPVI receptors lacking the PRD. These findings define a molecular priming mechanism that enables an immune-type receptor to adopt a hemostatic function. These studies also demonstrate that active kinases can constitutively associate with immune-type receptors without initiating signal transduction before receptor ligation, consistent with a recent molecular model of immune receptor signaling in which receptor ligation is required to bring active kinases to their receptor substrates.

[1]  Etienne Gagnon,et al.  Regulation of T Cell Receptor Activation by Dynamic Membrane Binding of the CD3ɛ Cytoplasmic Tyrosine-Based Motif , 2008, Cell.

[2]  G. Stoll,et al.  Molecular mechanisms of thrombus formation in ischemic stroke: novel insights and targets for treatment. , 2008, Blood.

[3]  R. Hynes,et al.  Structure-function analysis reveals discrete beta3 integrin inside-out and outside-in signaling pathways in platelets. , 2007, Blood.

[4]  T. Smithgall,et al.  HIV-1 Nef Selectively Activates Src Family Kinases Hck, Lyn, and c-Src through Direct SH3 Domain Interaction* , 2006, Journal of Biological Chemistry.

[5]  J. Ragoussis,et al.  The Leukocyte Receptor Complex in Chicken Is Characterized by Massive Expansion and Diversification of Immunoglobulin-Like Loci , 2006, PLoS genetics.

[6]  Michael Liss,et al.  Identification of preferred protein interactions by phage‐display of the human Src homology‐3 proteome , 2006, EMBO reports.

[7]  John P. Cannon,et al.  Reconstructing immune phylogeny: new perspectives , 2005, Nature Reviews Immunology.

[8]  D. Hammer,et al.  GPVI and α2β1 play independent critical roles during platelet adhesion and aggregate formation to collagen under flow , 2005 .

[9]  O. McCarty,et al.  GPVI and integrin αIIbβ3 signaling in platelets , 2005, Journal of thrombosis and haemostasis : JTH.

[10]  S. Watson,et al.  Delineation of the Region in the Glycoprotein VI Tail Required for Association with the Fc Receptor γ-Chain* , 2003, Journal of Biological Chemistry.

[11]  S. Watson,et al.  Platelet-collagen interaction: is GPVI the central receptor? , 2003, Blood.

[12]  Changdong Liu,et al.  Fc Rγ-independent Signaling by the Platelet Collagen Receptor Glycoprotein VI* , 2003, The Journal of Biological Chemistry.

[13]  S. Watson,et al.  Association of Fyn and Lyn with the Proline-rich Domain of Glycoprotein VI Regulates Intracellular Signaling* , 2002, The Journal of Biological Chemistry.

[14]  Y. Liu,et al.  The Platelet Receptor GPVI Mediates Both Adhesion and Signaling Responses to Collagen in a Receptor Density-dependent Fashion* , 2002, The Journal of Biological Chemistry.

[15]  B. Nieswandt,et al.  Glycoprotein VI but not α2β1 integrin is essential for platelet interaction with collagen , 2001 .

[16]  Giulio Superti-Furga,et al.  Dynamic Coupling between the SH2 and SH3 Domains of c-Src and Hck Underlies Their Inactivation by C-Terminal Tyrosine Phosphorylation , 2001, Cell.

[17]  S. Watson,et al.  Fyn and Lyn phosphorylate the Fc receptor gamma chain downstream of glycoprotein VI in murine platelets, and Lyn regulates a novel feedback pathway. , 2000, Blood.

[18]  T. Wells,et al.  The Platelet Collagen Receptor Glycoprotein VI Is a Member of the Immunoglobulin Superfamily Closely Related to FcαR and the Natural Killer Receptors* , 1999, The Journal of Biological Chemistry.

[19]  Brian Savage,et al.  Specific Synergy of Multiple Substrate–Receptor Interactions in Platelet Thrombus Formation under Flow , 1998, Cell.

[20]  H. Takayama,et al.  Physical and Functional Association of the Src Family Kinases Fyn and Lyn with the Collagen Receptor Glycoprotein VI-Fc Receptor γ Chain Complex on Human Platelets , 1998, The Journal of experimental medicine.

[21]  S. Watson,et al.  A Collagen-Like Peptide Stimulates Tyrosine Phosphorylation of syk and Phospholipase Cγ2 in Platelets Independent of the Integrin α2β1 , 1997 .

[22]  J. Kuriyan,et al.  Activation of the Sire-family tyrosine kinase Hck by SH3 domain displacement , 1997, Nature.

[23]  R. Rickles,et al.  Identification of Src, Fyn, Lyn, PI3K and Abl SH3 domain ligands using phage display libraries. , 1994, The EMBO journal.

[24]  M. Goldsmith,et al.  Early signal transduction by the antigen receptor without commitment to T cell activation. , 1988, Science.

[25]  Miriam K. Konkel,et al.  Tumour invasion and metastasis initiated by microRNA-10b in breast cancer , 2008, Nature.

[26]  S. Watson,et al.  University of Birmingham G6b-B inhibits constitutive and agonist-induced signaling by glycoprotein VI and CLEC-2 , 2008 .

[27]  D. Hammer,et al.  GPVI and alpha2beta1 play independent critical roles during platelet adhesion and aggregate formation to collagen under flow. , 2005, Blood.

[28]  B. Nieswandt,et al.  Glycoprotein VI but not alpha2beta1 integrin is essential for platelet interaction with collagen. , 2001, The EMBO journal.

[29]  Aldons J. Lusis,et al.  Atherosclerosis : Vascular biology , 2000 .

[30]  S. Watson,et al.  A collagen-like peptide stimulates tyrosine phosphorylation of syk and phospholipase C gamma2 in platelets independent of the integrin alpha2beta1. , 1997, Blood.