Dual ITAM-mediated proteolytic pathways for irreversible inactivation of platelet receptors: de-ITAM-izing Fc (cid:1) RIIa

CollagenbindingtoglycoproteinVI(GPVI) induces signals critical for platelet activation in thrombosis. Both ligand-induced GPVI signaling through its coassociated Fc-receptor (cid:1) -chain (FcR (cid:1) ) immuno-receptor tyrosine-activation motif (ITAM) and the calmodulin inhibitor, W7, dissoci-ate calmodulin from GPVI and induce metalloproteinase-mediated GPVI ectodomain shedding. We investigated whether signaling by another ITAM-bearing receptor on platelets, Fc (cid:1) RIIa, also down-regulates GPVI expression. Agonists that signal through Fc (cid:1) RIIa, the mAbs VM58 or 14A2, potently induced GPVI shedding, inhibitable by the metalloproteinase inhibitor, Unexpectedly, Fc (cid:1) RIIa also underwent rapid proteolysis in platelets treated with agonists for Fc (cid:1) RIIa (VM58/ 14A2) or GPVI/FcR (cid:1) (the snake toxin, convulxin), generating an approximate 30-kDa fragment. Immunoprecipitation/ pull-down experiments showed that Fc (cid:1) RIIa also bound calmodulin and W7 induced Fc (cid:1) RIIa cleavage. However, unlike GPVI, the approximate 30-kDa Fc (cid:1) RIIa fragment remained platelet associated, and proteolysis was unaffected by GM6001butwasinhibitedbyamembrane-permeable calpain inhibitor, E64d; consistent with this, (cid:2) -calpain cleaved an Fc (cid:1) RIIa tail-fusion protein at 222 Lys/ 223 Ala and 230 Gly/ 231 Arg, upstream of the ITAM domain. These findings suggest simulta-neous activation of distinct extracellular (metalloproteinase-mediated) and intracellular (calpain-mediated) proteolytic pathways irreversibly inactivating platelet GPVI/FcR (cid:1) and Fc (cid:1) RIIa, respectively. Activation of both pathways was observed with immunoglobulin from patients with heparin-induced thrombo-cytopenia (HIT), suggesting novel mechanisms for platelet dysfunction by Fc (cid:1) RIIa after immunologic insult. (Blood. 2008; 111:165-174)

[1]  M. Berndt,et al.  Controlled shedding of platelet glycoprotein (GP)VI and GPIb–IX–V by ADAM family metalloproteinases , 2007, Journal of thrombosis and haemostasis : JTH.

[2]  C. Gachet,et al.  Calmodulin interacts with the platelet ADP receptor P2Y1. , 2006, The Biochemical journal.

[3]  E. Kremmer,et al.  Expression of platelet collagen receptor glycoprotein VI is associated with acute coronary syndrome. , 2006, European heart journal.

[4]  Ingemar Lundström,et al.  Phosphorylation of Thr654 but not Thr669 within the juxtamembrane domain of the EGF receptor inhibits calmodulin binding. , 2006, Biochemical and biophysical research communications.

[5]  P. Newman,et al.  Activation-independent, antibody-mediated removal of GPVI from circulating human platelets: development of a novel NOD/SCID mouse model to evaluate the in vivo effectiveness of anti-human platelet agents. , 2006, Blood.

[6]  H. Trist,et al.  Raft localisation of FcgammaRIIa and efficient signaling are dependent on palmitoylation of cysteine 208. , 2006, Immunology letters.

[7]  C. Balduini,et al.  A new role for FcγRIIA in the potentiation of human platelet activation induced by weak stimulation , 2005 .

[8]  M. Kinter,et al.  Calmodulin Binds to the Cytoplasmic Domain of Angiotensin-converting Enzyme and Regulates Its Phosphorylation and Cleavage Secretion* , 2005, Journal of Biological Chemistry.

[9]  M. Berndt,et al.  Role of calmodulin in platelet receptor function. , 2005, Current medicinal chemistry. Cardiovascular and hematological agents.

[10]  Anna Huttenlocher,et al.  Regulating cell migration: calpains make the cut , 2005, Journal of Cell Science.

[11]  M. Pelto-huikko,et al.  Shedding light on ADAM metalloproteinases. , 2005, Trends in biochemical sciences.

[12]  Xu Feng.  RANKing Intracellular Signaling in Osteoclasts , 2005, IUBMB life.

[13]  Eric Vivier,et al.  KARAP/DAP12/TYROBP: three names and a multiplicity of biological functions , 2005, European journal of immunology.

[14]  B. Nieswandt,et al.  Evidence for a Role of ADAM17 (TACE) in the Regulation of Platelet Glycoprotein V* , 2005, Journal of Biological Chemistry.

[15]  Robert K Andrews,et al.  Regulation of platelet membrane levels of glycoprotein VI by a platelet-derived metalloproteinase. , 2004, Blood.

[16]  Y. Ozaki,et al.  Glycoprotein VI is associated with GPIb-IX-V on the membrane of resting and activated platelets , 2004, Thrombosis and Haemostasis.

[17]  Y. Yatomi,et al.  Cleavage of platelet endothelial cell adhesion molecule‐1 (PECAM‐1) in platelets exposed to high shear stress , 2004, Journal of thrombosis and haemostasis : JTH.

[18]  Ya A Naimushin,et al.  Von Willebrand factor can support platelet aggregation via interaction with activated GPIIb–IIIa and GPIb , 2004, Platelets.

[19]  S. Jackson,et al.  Platelet Factor XIII and Calpain Negatively Regulate Integrin αIIbβ3 Adhesive Function and Thrombus Growth* , 2004, Journal of Biological Chemistry.

[20]  Lai-Man Lau,et al.  Proteolytic cleavage of platelet endothelial cell adhesion molecule‐1 (PECAM‐1/CD31) is regulated by a calmodulin‐binding motif , 2004, FEBS letters.

[21]  Peter Tompa,et al.  On the Sequential Determinants of Calpain Cleavage* , 2004, Journal of Biological Chemistry.

[22]  D. Wagner,et al.  GPVI down-regulation in murine platelets through metalloproteinase-dependent shedding , 2004, Thrombosis and Haemostasis.

[23]  J. Hartwig,et al.  Metalloproteinase inhibitors improve the recovery and hemostatic function of in vitro-aged or -injured mouse platelets. , 2003, Blood.

[24]  Lisa A. Pitcher,et al.  T-cell receptor signal transmission: who gives an ITAM? , 2003, Trends in immunology.

[25]  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.

[26]  J. Whisstock,et al.  Glycoprotein Ib-IX-V. , 2003, The international journal of biochemistry & cell biology.

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

[28]  D. E. Goll,et al.  The calpain system. , 2003, Physiological reviews.

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

[30]  S. Groshen,et al.  Increased platelet Fc receptor expression as a potential contributing cause of platelet hypersensitivity to collagen in diabetes mellitus , 2003, British journal of haematology.

[31]  R. Bodnar,et al.  Regulation of Glycoprotein Ib-IX-von Willebrand Factor Interaction by cAMP-dependent Protein Kinase-mediated Phosphorylation at Ser 166 of Glycoprotein Ibβ* , 2002, The Journal of Biological Chemistry.

[32]  S. Groshen,et al.  Potential role of platelet FcgammaRIIA in collagen-mediated platelet activation associated with atherothrombosis. , 2002, Atherosclerosis.

[33]  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.

[34]  S. Watson,et al.  Interaction of calmodulin with the cytoplasmic domain of platelet glycoprotein VI. , 2002, Blood.

[35]  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.

[36]  C. Mitchell,et al.  Interaction of calmodulin with the cytoplasmic domain of the platelet membrane glycoprotein Ib-IX-V complex. , 2001, Blood.

[37]  J. Fox,et al.  Cytoskeletal Proteins and Platelet Signaling , 2001, Thrombosis and Haemostasis.

[38]  D. Burshtyn,et al.  Intracellular Signaling by the Killer Immunoglobulin-Like Receptors and Ly49 , 2001, Science's STKE.

[39]  B. Nieswandt,et al.  Long-Term Antithrombotic Protection by in Vivo Depletion of Platelet Glycoprotein VI in Mice , 2001, The Journal of experimental medicine.

[40]  Xiaoping Du,et al.  Evidence That β3 Integrin-Induced Rac Activation Involves the Calpain-Dependent Formation of Integrin Clusters That Are Distinct from the Focal Complexes and Focal Adhesions That Form as Rac and Rhoa Become Active , 2000, The Journal of cell biology.

[41]  W. Vainchenker,et al.  Cloning, characterization, and functional studies of human and mouse glycoprotein VI: a platelet-specific collagen receptor from the immunoglobulin superfamily. , 2000, Blood.

[42]  B. Nieswandt,et al.  Expression and Function of the Mouse Collagen Receptor Glycoprotein VI Is Strictly Dependent on Its Association with the FcRγ Chain* , 2000, The Journal of Biological Chemistry.

[43]  J. Whisstock,et al.  Requirement of leucine-rich repeats of glycoprotein (GP) Ibalpha for shear-dependent and static binding of von Willebrand factor to the platelet membrane GP Ib-IX-V complex. , 2000, Blood.

[44]  R. Bodnar,et al.  The Cytoplasmic Domain of the Platelet Glycoprotein Ibα Is Phosphorylated at Serine 609* , 1999, The Journal of Biological Chemistry.

[45]  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.

[46]  Scott M. Taylor,et al.  The role of the human Fc receptor Fc gamma RIIA in the immune clearance of platelets: a transgenic mouse model. , 1999, Journal of immunology.

[47]  M. Jutila,et al.  Calmodulin Regulates L-Selectin Adhesion Molecule Expression and Function through a Protease-Dependent Mechanism , 1998, Cell.

[48]  W. Foss,et al.  Physical Proximity and Functional Interplay of the Glycoprotein Ib-IX-V Complex and the Fc Receptor FcγRIIA on the Platelet Plasma Membrane* , 1998, The Journal of Biological Chemistry.

[49]  M. Swindells,et al.  Solution structure of calmodulin-W-7 complex: the basis of diversity in molecular recognition. , 1998, Journal of molecular biology.

[50]  S. Harris,et al.  Binding of purified 14-3-3 zeta signaling protein to discrete amino acid sequences within the cytoplasmic domain of the platelet membrane glycoprotein Ib-IX-V complex. , 1998, Biochemistry.

[51]  J. Martín-Nieto,et al.  The human epidermal growth factor receptor contains a juxtamembrane calmodulin-binding site. , 1998, Biochemistry.

[52]  S. Watson,et al.  Glycoprotein VI is the collagen receptor in platelets which underlies tyrosine phosphorylation of the Fc receptor γ‐chain , 1997 .

[53]  T. Wells,et al.  Platelet Activation and Signal Transduction by Convulxin, a C-type Lectin from Crotalus durissus terrificus (Tropical Rattlesnake) Venom via the p62/GPVI Collagen Receptor* , 1997, The Journal of Biological Chemistry.

[54]  S. Jackson,et al.  Calpain Cleavage of Focal Adhesion Proteins Regulates the Cytoskeletal Attachment of Integrin αIIbβ3 (Platelet Glycoprotein IIb/IIIa) and the Cellular Retraction of Fibrin Clots* , 1997, The Journal of Biological Chemistry.

[55]  J. J. Roberts,et al.  Platelet activation induced by a murine monoclonal antibody directed against a novel tetra‐span antigen , 1995, British journal of haematology.

[56]  M. Berndt,et al.  Detection of an epitope specific for the dissociated form of glycoprotein IIIa of platelet membrane glycoprotein IIb‐IIIa complex and its expression on the surface of adherent platelets , 1993, British journal of haematology.

[57]  J. Hoxie,et al.  Activation of Fc gamma RII induces tyrosine phosphorylation of multiple proteins including Fc gamma RII. , 1992, The Journal of biological chemistry.

[58]  L. Ashman,et al.  The murine monoclonal antibody, 14A2.H1, identifies a novel platelet surface antigen , 1991, British journal of haematology.

[59]  G. Demartino,et al.  Calcium-activated neutral protease (calpain) system: structure, function, and regulation. , 1991, Physiological reviews.

[60]  V. Makarov,et al.  [Inhibition of Fc-receptor dependent platelet aggregation by monoclonal antibodies against the glycoprotein IIb-IIIa complex]. , 1991, Biokhimiia.

[61]  Lubowski Tj.  Heparin-induced thrombocytopenia and thrombosis. , 1990, Connecticut medicine.

[62]  M. Moroi,et al.  A patient with platelets deficient in glycoprotein VI that lack both collagen-induced aggregation and adhesion. , 1989, The Journal of clinical investigation.

[63]  J. Gorman,et al.  Purification of botrocetin from Bothrops jararaca venom. Analysis of the botrocetin-mediated interaction between von Willebrand factor and the human platelet membrane glycoprotein Ib-IX complex. , 1989, Biochemistry.

[64]  K. Wang,et al.  Calmodulin-binding proteins as calpain substrates. , 1989, The Biochemical journal.

[65]  M. Hibbs,et al.  Molecular cloning of a human immunoglobulin G Fc receptor. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[66]  M. Berndt,et al.  Ristocetin-dependent reconstitution of binding of von Willebrand factor to purified human platelet membrane glycoprotein Ib-IX complex. , 1988, Biochemistry.

[67]  C. Anderson,et al.  Identification of a second class of IgG Fc receptors on human neutrophils. A 40 kilodalton molecule also found on eosinophils , 1986, The Journal of experimental medicine.

[68]  H. Zola,et al.  Molecular characterization of quinine/quinidine drug-dependent antibody platelet interaction using monoclonal antibodies. , 1985, Blood.

[69]  H. Zola,et al.  Purification and preliminary characterization of the glycoprotein Ib complex in the human platelet membrane. , 1985, European journal of biochemistry.

[70]  J. Ravetch,et al.  Fcgamma receptors: old friends and new family members. , 2006, Immunity.

[71]  J. Villeval,et al.  Platelet activation induces metalloproteinase-dependent GP VI cleavage to down-regulate platelet reactivity to collagen. , 2005, Blood.

[72]  P. Barton,et al.  Biochemical analysis and crystallisation of Fc gamma RIIa, the low affinity receptor for IgG. , 1999, Immunology letters.

[73]  P. Barton,et al.  Crystal structure of the human leukocyte Fc receptor, Fc gammaRIIa. , 1999, Nature structural biology.

[74]  R. Lindeman,et al.  Single amino acid mutation of Fc gamma receptor is associated with the development of heparin-induced thrombocytopenia. , 1995, British journal of haematology.

[75]  M. Hulett,et al.  Molecular basis of Fc receptor function. , 1994, Advances in immunology.

[76]  D. E. Goll,et al.  Immunolocalization of the calpains and calpastatin in human and bovine platelets. , 1991, Biomedica biochimica acta.

[77]  T. Tanaka,et al.  Calcium-regulated modulator protein interacting agents inhibit smooth muscle calcium-stimulated protein kinase and ATPase. , 1980, Molecular pharmacology.

[78]  The Journal of Experimental Medicine A Crucial Role of Glycoprotein VI for Platelet Recruitment to the Injured Arterial Wall In Vivo , 2022 .