Recruitment and Activation of SHP-1 Protein-tyrosine Phosphatase by Human Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1)

Stimulation of platelet aggregation leads to tyrosine phosphorylation of a number of receptors and signaling molecules including platelet endothelial cell adhesion molecule-1 (PECAM-1). In this report, we demonstrate that both protein-tyrosine phosphatases SHP-1 and SHP-2 physically associate with different kinetics of assembly with tyrosine-phosphorylated human PECAM-1 during integrin αIIbβ3-mediated platelet aggregation. Peptido-precipitation analysis revealed that tyrosine-phosphorylated peptides encompassing residues 658–668 and 681–691 of PECAM-1 bound specifically to both protein-tyrosine phosphatases SHP-1 and SHP-2. We further show that the association of SHP-1 with PECAM-1 occurs through the direct interaction of thesrc homology region 2 domains of SHP-1 with two highly conserved phosphotyrosine binding motifs within PECAM-1 having the sequences NSDVQpY663TEVQV and DTETVpY686SEVRK (where pY represents phosphotyrosine). In vitrodephosphorylation experiments using phosphotyrosyl PECAM-1 peptides encompassing either Tyr-663 or Tyr-686 revealed induction of SHP-1 catalytic activity, suggesting that PECAM-1 serves as a SHP-1 substrate. Surface plasmon resonance studies reveal that recombinant SHP-2 binds PECAM-1 phosphopeptides with 5-fold higher affinity than recombinant SHP-1. These data suggest that in hematopoietic cells such as platelets, PECAM-1 cellular signaling is regulated by the selective recruitment and activation of two distinct protein-tyrosine phosphatases, SHP-1 and SHP-2, via a common immunoreceptor tyrosine-based inhibitory-like motif. Stimulation of platelet aggregation leads to tyrosine phosphorylation of a number of receptors and signaling molecules including platelet endothelial cell adhesion molecule-1 (PECAM-1). In this report, we demonstrate that both protein-tyrosine phosphatases SHP-1 and SHP-2 physically associate with different kinetics of assembly with tyrosine-phosphorylated human PECAM-1 during integrin αIIbβ3-mediated platelet aggregation. Peptido-precipitation analysis revealed that tyrosine-phosphorylated peptides encompassing residues 658–668 and 681–691 of PECAM-1 bound specifically to both protein-tyrosine phosphatases SHP-1 and SHP-2. We further show that the association of SHP-1 with PECAM-1 occurs through the direct interaction of thesrc homology region 2 domains of SHP-1 with two highly conserved phosphotyrosine binding motifs within PECAM-1 having the sequences NSDVQpY663TEVQV and DTETVpY686SEVRK (where pY represents phosphotyrosine). In vitrodephosphorylation experiments using phosphotyrosyl PECAM-1 peptides encompassing either Tyr-663 or Tyr-686 revealed induction of SHP-1 catalytic activity, suggesting that PECAM-1 serves as a SHP-1 substrate. Surface plasmon resonance studies reveal that recombinant SHP-2 binds PECAM-1 phosphopeptides with 5-fold higher affinity than recombinant SHP-1. These data suggest that in hematopoietic cells such as platelets, PECAM-1 cellular signaling is regulated by the selective recruitment and activation of two distinct protein-tyrosine phosphatases, SHP-1 and SHP-2, via a common immunoreceptor tyrosine-based inhibitory-like motif. focal adhesion kinase thrombin receptor-activating peptide B cell IgG Fc receptor IgGFcγRIIIa receptor IgG FcγRIIa receptor β-subunit of the high affinity IgE receptor src homology 2 protein-tyrosine phosphatase NH2-terminal SH2 domain of SHP-1 COOH-terminal SH2 domain of SHP-1 immunoreceptor tyrosine-based activation motif immunoreceptor tyrosine-based inhibitory motif glutathioneS-transferase polyacrylamide gel electrophoresis N, N, N′,N′-tetramethylethylenediamine platelet endothelial cell adhesion molecule-1. Platelets possess a repertoire of cell adhesion receptors, including the major fibrinogen receptor, integrin αIIbβ3 (GPIIb-IIIa), which play a pivotal role in platelet cell adhesion, aggregation, and clot retraction (for a review, see Ref. 1Hynes R.O. Cell. 1992; 69: 11-25Abstract Full Text PDF PubMed Scopus (8941) Google Scholar). Engagement of αIIbβ3converts the integrin to its active conformation, making it accessible for binding its soluble ligand, fibrinogen (2Gartner T.K. Bennett J.S. J. Biol. Chem. 1985; 260: 11891-11894Abstract Full Text PDF PubMed Google Scholar, 3Plow E.F. Pierschbacher M.D. Ruoslahti E. Marguerie G.A. Ginsberg M.H. Proc. Natl. Acad. Sci. U. S. A. 1985; 82: 8057-8061Crossref PubMed Scopus (441) Google Scholar). This event is followed by downstream clustering that triggers cytoskeletal reorganization and activation of protein-tyrosine kinases (4Shattil S.J. Haimovich B. Cunningham M. Lipfert L. Parsons J.T. Ginsberg M.H. Brugge J.S. J. Biol. Chem. 1994; 269: 14738-14745Abstract Full Text PDF PubMed Google Scholar, 5Golden A. Brugge J.S. Shattil S.J. J. Cell Biol. 1990; 111: 3117-3127Crossref PubMed Scopus (184) Google Scholar). Platelets offer an attractive model to study protein tyrosine phosphorylation, since the early and late waves of tyrosine phosphorylation can be dissected by examining the effects of agonist-induced cellular activation followed by platelet/platelet cell contact initiated by the stirring of platelets (aggregation). Integrin engagement and aggregation of platelets results in tyrosine phosphorylation of a number of receptors and signaling molecules as a late event, including pp125FAK,1pp60c-src, integrin β3, and platelet endothelial cell adhesion molecule-1 (PECAM-1) (5Golden A. Brugge J.S. Shattil S.J. J. Cell Biol. 1990; 111: 3117-3127Crossref PubMed Scopus (184) Google Scholar, 6Ferrell Jr., J.E. Martin G.S. Proc. Natl. Acad. Sci. U. S. A. 1989; 86: 2234-2238Crossref PubMed Scopus (204) Google Scholar, 7Law D.A. Nannizzi-Alaimo L. Phillips D.R. J. Biol. Chem. 1996; 271: 10811-10815Abstract Full Text Full Text PDF PubMed Scopus (204) Google Scholar, 8Lipfert L. Haimovich B. Schaller M.D. Cobb B.S. Parson J.T. Brugge J.S. J. Cell Biol. 1992; 119: 905-912Crossref PubMed Scopus (624) Google Scholar, 9Jackson D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar). PECAM-1 is a 130-kDa transmembrane glycoprotein that is expressed predominantly on the surface of platelets, monocytes, neutrophils, and selected T-lymphocyte subsets and is concentrated at the apical borders of endothelial cells (10Newman P.J. Berndt M.C. Gorski J. White G.C. Lyman S. Paddock C. Muller W.A. Science. 1990; 247: 1219-1222Crossref PubMed Scopus (813) Google Scholar). Its structural organization consists of six extracellular Ig-like homology domains, a transmembrane domain, and a cytoplasmic domain (10Newman P.J. Berndt M.C. Gorski J. White G.C. Lyman S. Paddock C. Muller W.A. Science. 1990; 247: 1219-1222Crossref PubMed Scopus (813) Google Scholar, 11Kirschbaum N.E. Gumina R.J. Newman P.J. Blood. 1994; 84: 4028-4037Crossref PubMed Google Scholar). Recent studies have suggested that PECAM-1 plays a role in outside-in signal transduction by transmitting signals into the cell interior resulting in its tyrosine phosphorylation and subsequent up-regulation of integrin function (12Varon D. Jackson D.E. Shenkman B. Dardik R. Tamarin I. Savion N. Newman P.J. Blood. 1998; 91: 500-508Crossref PubMed Google Scholar, 13Leavesley D.I. Oliver J.M. Swart B.W. Berndt M.C. Haylock D.N. Simmons P.J. J. Immunol. 1994; 153: 4673-4683PubMed Google Scholar, 14Tanaka Y. Albelda S.M. Horgan K.J. Van Seventer G.A. Shimizu Y. Newman W. Hallam J. Newman P.J. Buck C.A. Shaw S. J. Exp. Med. 1992; 176: 245-253Crossref PubMed Scopus (337) Google Scholar, 15Berman M.E. Muller W.A. J. Immunol. 1995; 154: 299-307PubMed Google Scholar). Since PECAM-1 lacks intrinsic tyrosine kinase activity, tyrosine phosphorylation of its cytoplasmic domain after its engagement is achieved through recruitment of cytosolic protein-tyrosine kinases such as pp60c-src (16Osawa M. Masuda M. Harada N. Lopes J. Cell Biol. 1997; Google Scholar, M. S. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). of tyrosine residues by the of protein-tyrosine kinases for recruitment and activation of src homology 2 cytosolic signaling of cytosolic molecules the expressed protein-tyrosine SHP-2 as and Recent studies have the of SHP-2, as by in and hematopoietic in cells a selected of the SH2 domain of SHP-2 M. S. R. J. 1997; PubMed Scopus Google Scholar, R. Cell. Biol. 1997; PubMed Scopus Google Scholar). In to binding receptors, SHP-2 binds to tyrosine-phosphorylated PECAM-1 at phosphotyrosine residues and via of its SH2 domains, which results in induction of catalytic of the phosphatase D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar, D.E. Newman P.J. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar, M. M. Harada N. 1997; PubMed Scopus Google Scholar, M. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). SHP-2 structural with protein-tyrosine SHP-1. phosphatases two SH2 domains, a catalytic phosphatase domain and a COOH-terminal region two tyrosine phosphorylation to have distinct on the of signaling Since SHP-1 a we that the phosphotyrosine binding motifs in the cytoplasmic domain of PECAM-1 be of with SHP-1 protein-tyrosine phosphatase in to SHP-2. SHP-1 as and is a cytosolic protein-tyrosine phosphatase that is expressed in hematopoietic cells is in cell J. Proc. Natl. Acad. Sci. U. S. A. 1992; PubMed Scopus Google Scholar). of the SHP-1 in results in a by hematopoietic and R.J. D.R. Cell. Full Text PDF PubMed Scopus Google Scholar, L. PubMed Scopus Google Scholar, L. Exp. 1989; Google Scholar). SHP-1 to associate with and receptors, including receptor, and U. U. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar, J. Blood. 1995; PubMed Google Scholar, D. I. Science. 1995; PubMed Scopus Google Scholar, L. A. D. M. R. A. L. E. J. Immunol. 1996; Google Scholar, R.J. J. Science. 1995; 269: PubMed Scopus Google Scholar, S. U. R. A. E. A. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar, Y. A. N. M. M. Cell. Biol. 1996; PubMed Scopus Google Scholar). interaction of SHP-1 with receptors via its SH2 domains to play a role in cellular signaling D. I. Science. 1995; PubMed Scopus Google Scholar, R.J. J. Science. 1995; 269: PubMed Scopus Google Scholar, 1995; Full Text PDF PubMed Scopus Google Scholar). SHP-1 to function in the and of and B cell and distinct with SHP-2 (for a review, see Ref. Cell. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). the of protein-tyrosine phosphatases in platelet cell and signaling of be to the role of phosphatases in adhesion receptors with signaling of this to the and for PECAM-1 to and two distinct protein-tyrosine phosphatases, SHP-1 and SHP-2. Since SHP-1 and SHP-2 protein-tyrosine phosphatases both expressed in hematopoietic human platelets as a model to study the kinetics of assembly during agonist-induced cellular activation and We show that tyrosine-phosphorylated PECAM-1 physically with both SHP-1 and SHP-2 with kinetics during integrin αIIbβ3-mediated platelet aggregation. interaction of SHP-1 and SHP-2 with PECAM-1 occurs through the direct interaction of the SH2 domains of SHP-1 and SHP-2 with two highly conserved phosphotyrosine binding motifs within PECAM-1 having the sequences and (where pY represents phosphotyrosine). These two phosphotyrosine binding Tyr-663 and Tyr-686 of catalytic activation of SHP-1. results suggest that cellular signaling in the of hematopoietic cells is regulated by the selective recruitment and activation of SHP-1 and SHP-2 via a common immunoreceptor tyrosine-based inhibitory motif motif of and and and platelet by to the PECAM-1 for the homology domain and PECAM-1 by Newman of These have 1995; Full Text PDF PubMed Scopus Google Scholar, Cell. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). of to the and COOH-terminal SH2 domains of SHP-1 SHP-2 and SHP-1 to the and COOH-terminal SH2 domains of SHP-2 to SHP-2 and SHP-1 These specifically protein-tyrosine SHP-2 and as a a platelet cell of IgG and IgG Jackson Platelets within the platelets to D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar, P.J. C.A. R. Berndt M.C. J. J. Cell Biol. 1992; 119: PubMed Scopus Google Scholar). of in the of 2 and fibrinogen with the with for with stirring for the and peptide to the of with stirring at for in a aggregation Platelets that by activation or aggregation in of and 2 and on a at for and by at for at as P.J. C.A. R. Berndt M.C. J. J. Cell Biol. 1992; 119: PubMed Scopus Google Scholar). of the the with of a of for at and at for cell with either or at as D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar). the by for at in of and to and by or to SHP-2, or PECAM-1 In the the with a IgG by using an of the of different by a which the in and PECAM-1 peptides with or encompassing of the tyrosine residues within the cytoplasmic domain of human PECAM-1 as D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar). the phosphopeptides in this human PECAM-1 phosphopeptides have to and the of the tyrosine residues including the signal of human PECAM-1 of tyrosine in of of the tyrosine residues is on the of the human PECAM-1 the signal pY tyrosine that the with an at to in a of the tyrosine residues is on the of the human PECAM-1 the signal pY tyrosine that the with an at to of PECAM-1 peptide with or with 2 of platelet at with of to the peptide and for a further at with with and the by for in of and by using SHP-1 SHP-2 at a of followed by the of a IgG at a of and SHP-1 and SHP-2 and and COOH-terminal SH2 domains of either SHP-1 or SHP-2 into by encompassing and SHP-1 or and SHP-2 or the SH2 domains of either SHP-1 or SHP-2 by with and using as U. M. I. J. Biol. Chem. 1994; 269: Full Text PDF PubMed Google Scholar). recombinant to be than on a of PECAM-1 peptides with or with encompassing the SH2 domains of either SHP-1 or SHP-2 D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar). by in and by of recombinant protein encompassing the SH2 domains of SHP-1 or SHP-2 by with to the and SH2 domains of either SHP-1 or SHP-2. of SHP-1 at in a as PECAM-1 peptide and of recombinant protein SHP-1 by thrombin during a at the with of and the of the at experiments that of surface plasmon resonance on a of PECAM-1 phosphopeptides a through of accessible by binding of a resonance of binding and of have D.E. Newman P.J. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). binding a of for recombinant SHP-1 and SHP-2 and to be the PECAM-1 phosphopeptides using and using to using as binding data to with on and studies have that two 658–668 and in the cytoplasmic domain of human PECAM-1 for the association of the protein-tyrosine SHP-2, Tyr-663 and Tyr-686 to for an SH2 interaction D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar, M. S. R. J. 1997; PubMed Scopus Google Scholar). Since SHP-2 structural with cytosolic protein-tyrosine SHP-1 and SHP-2 the and SHP-1 we that the tyrosine phosphorylation and in the PECAM-1 cytoplasmic domain associate with this cytosolic signaling this we a of PECAM-1 cytoplasmic domain peptides with or to of the PECAM-1 receptor to platelet using and the of SHP-1 and SHP-2 by with SHP-1 and SHP-2 in analysis using a SHP-2 revealed that two PECAM-1 phosphopeptides and bound the protein-tyrosine SHP-2, the peptides tyrosine-phosphorylated at Tyr-663 and we a with the PECAM-1 peptides with or as in and with a SHP-1 we a SHP-1 binding to the PECAM-1 phosphopeptides 658–668 and 681–691 of SHP-1 with of the two peptides since of the be by a of PECAM-1 phosphopeptides and These data suggest that PECAM-1 can associate with two distinct protein-tyrosine phosphatases, SHP-2 and via common tyrosine binding motifs in the cytoplasmic domain of SHP-1 can associate with and cell surface receptors including receptor receptor, receptor, high affinity IgE receptor and via its SH2 domains U. U. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar, J. Blood. 1995; PubMed Google Scholar, D. I. Science. 1995; PubMed Scopus Google Scholar, L. A. D. M. R. A. L. E. J. Immunol. 1996; Google Scholar, R.J. J. Science. 1995; 269: PubMed Scopus Google Scholar, S. U. R. A. E. A. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar, Y. A. N. M. M. Cell. Biol. 1996; PubMed Scopus Google Scholar). the SH2 domains of SHP-1 the interaction with we recombinant encompassing the and SH2 domains of SHP-1 and the of the PECAM-1 phosphopeptides 658–668 and 681–691 in the and of at Tyr-663 and Tyr-686 to to SH2 domains of SHP-1. with and the of the SH2 domains of SHP-1 using a the and SH2 domains of SHP-1. in 2 the PECAM-1 peptides to associate with either and or the 2 and In the PECAM-1 phosphopeptides 658–668 and 681–691 encompassing Tyr-663 and Tyr-686 bound to a protein the and SH2 domains of SHP-1 and This interaction since the PECAM-1 phosphopeptides to associate with and results with PECAM-1 phosphopeptides binding to a protein the and SH2 domains of SHP-2 and as we have D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google 2 These data demonstrate that the interaction of SHP-1 with tyrosine-phosphorylated PECAM-1 cytoplasmic domain peptides is by the and SH2 domains of SHP-1. studies have that PECAM-1 tyrosine-phosphorylated during integrin αIIbβ3-mediated platelet aggregation D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google Scholar). the association of tyrosine-phosphorylated PECAM-1 with SHP-1 suggested by the PECAM-1 binding studies in human platelets, we PECAM-1 of platelets that by by by platelets, and by of platelet aggregation by peptide to and by analysis for the of SHP-1. in a of SHP-1 with PECAM-1 platelets by this association is platelet aggregation is to This interaction is and tyrosine phosphorylation of since with peptide the We the kinetics of assembly of by studies with at and stirring the platelets in the for the to the platelets the to with and the by analysis with and SHP-1 and SHP-2 can be in the of PECAM-1 is at a of in the of platelet aggregation, tyrosine phosphorylation of PECAM-1 is by and and after tyrosine phosphorylation of PECAM-1 with the association of two tyrosine-phosphorylated with a of that to SHP-1 and the with a of that to be and the with an the of SHP-1 SHP-2 to the that its by is and that it is tyrosine-phosphorylated during integrin αIIbβ3-mediated platelet aggregation. This is in to which is tyrosine-phosphorylated platelet aggregation with an suggested that SHP-2 association with PECAM-1 the tyrosine phosphorylation of PECAM-1 and occurs with different kinetics of assembly than SHP-1 of association with PECAM-1 to be of SHP-1 and of SHP-2 of PECAM-1 by analysis These data suggest that tyrosine-phosphorylated cellular PECAM-1 can with two distinct protein-tyrosine phosphatases during platelet aggregation. that SHP-1 is tyrosine-phosphorylated during platelet aggregation different of phosphatases in cellular signaling in of assembly of association with tyrosine-phosphorylated cellular PECAM-1 during integrin αIIbβ3-mediated platelet platelets with either or with and to in the of 2 and the with on and to analysis with a of and with an a or a to SHP-2 that of a tyrosine-phosphorylated to SHP-1 occurs with tyrosine phosphorylation of PECAM-1 at to of platelet aggregation. of and expressed in and the of SHP-2, and the of IgG protein-tyrosine SHP-2, tyrosine-phosphorylated integrin αIIbβ3-mediated platelet aggregation. phosphotyrosine of SHP-1 the of platelets platelets and by peptide to and with to show of SHP-1 into phosphotyrosine of SHP-2 the of platelets platelets and of peptide to and with to show of SHP-2 into SHP-1 is tyrosine-phosphorylated platelet studies have that protein-tyrosine phosphatases such as SHP-1 in a in of SH2 domains by to phosphotyrosine binding motifs leads to of the inhibitory of the SH2 domain on its phosphatase domain and results in an of the phosphatase and induction of catalytic activity, a that is further by its tyrosine phosphorylation S. J. Biol. Chem. 1994; 269: Full Text PDF PubMed Google Scholar). Recent data on SHP-2 have revealed that the catalytic domain is regulated by its domain as a in catalytic its domain to binding and S. S. Cell. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar). Since studies suggest that PECAM-1 cytoplasmic domain 658–668 Tyr-663 and 681–691 Tyr-686 phosphopeptides to with SHP-1 via its SH2 domains, we that the association to induction of the catalytic activation of SHP-1. this we the tyrosine-phosphorylated peptides to the tyrosine phosphorylation to be in the cytoplasmic domain of PECAM-1 to the of SH2 domain on SHP-1 catalytic in using a two of the phosphopeptides Tyr-663 and a in activation of SHP-1 phosphatase of SHP-1 by PECAM-1 phosphopeptides Tyr-663 and Tyr-686 a activation of on the of the PECAM-1 tyrosine binding we that Tyr-663 having the in the to the phosphotyrosine a higher catalytic activation of SHP-1. we a higher in SHP-1 activation for Tyr-663 with a with the Tyr-686 PECAM-1 in the of the of peptides on the catalytic of SHP-1 the data suggest that the of the SH2 domains of SHP-1 by PECAM-1 tyrosine binding motifs Tyr-663 and Tyr-686 leads to induction of catalytic activation of the the that SHP-1 and SHP-2 to phosphotyrosine residues and in PECAM-1 in an SH2 we the affinity for the interaction of recombinant of SHP-1 and SHP-2 with and by surface plasmon for the binding of and to and peptides in PECAM-1 bound to with a of of PECAM-1 bound to with a of In PECAM-1 bound to with a of PECAM-1 bound to with of In of we that the binding of recombinant the by surface plasmon binding using recombinant with of PECAM-1 peptides results suggest that SHP-2 binds the motif in PECAM-1 with at 5-fold higher affinity than for the binding of recombinant SHP-1 and SHP-2 with human PECAM-1 PECAM-1 analysis of a of recombinant binding by surface plasmon resonance at the after binding of the recombinant pY tyrosine in a analysis of a of recombinant binding by surface plasmon resonance at the after binding of the recombinant pY tyrosine studies have that PECAM-1 tyrosine-phosphorylated in to integrin engagement on extracellular M. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google integrin αIIbβ3-mediated platelet aggregation D.E. Ward C.M. Wang R. Newman P.J. J. Biol. Chem. 1997; 272: 6986-6993Abstract Full Text Full Text PDF PubMed Scopus (198) Google (16Osawa M. Masuda M. Harada N. Lopes J. Cell Biol. 1997; Google aggregation of high affinity IgE receptor W. J. E. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google and by (12Varon D. Jackson D.E. Shenkman B. Dardik R. Tamarin I. Savion N. Newman P.J. Blood. 1998; 91: 500-508Crossref PubMed Google for cytosolic signaling molecules such as SHP-2. SHP-2 structural homology with SHP-1 protein-tyrosine These protein-tyrosine phosphatases of SH2 domains, a catalytic domain, and COOH-terminal tyrosine which can as for molecules such as of within SH2 domains and for binding peptide residues to binding motifs of the sequences M. S. R.J. J.E. B. Cell. Full Text PDF PubMed Scopus Google Scholar). SHP-2 binds SHP-1 binds of the PECAM-1 cytoplasmic domain and its with SHP-1 and SHP-2 binding that the sequences and the for binding or both protein-tyrosine phosphatases, SHP-1 and SHP-2. at the to the phosphotyrosine a higher affinity binding for SHP-2 than since SHP-1 a at this In analysis of tyrosine-phosphorylated and peptides revealed that two of PECAM-1 with distinct of two protein-tyrosine phosphatases, SHP-1 and SHP-2, in a and results that PECAM-1 with SHP-1 and SHP-2 of integrin αIIbβ3-mediated platelet aggregation and of the and affinity of and that SHP-2 associate with higher affinity than SHP-1 within the and of SHP-1 association with tyrosine-phosphorylated cellular PECAM-1 to the during platelet aggregation. tyrosine phosphorylation within PECAM-1 by residues an by a that in of an receptor such as inhibitory receptor, and E. J. Immunol. 1997; Google Scholar, E. M. Immunol. 1997; Full Text PDF PubMed Scopus Google Scholar, Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar). motifs have a tyrosine residues to binding of protein-tyrosine phosphatases such as SHP-2, the with immunoreceptor tyrosine-based activation motif receptors, resulting in phosphorylation by kinases and recruitment of which results in inhibitory signals to receptor signaling S. L. A. J.E. L. M. E. J. Immunol. 1997; PubMed Scopus Google J. Exp. Med. 1995; PubMed Scopus Google Scholar). receptors have as phosphorylation of tyrosine and and tyrosine kinases M. 1989; PubMed Scopus Google Scholar). of receptors and receptors such as receptor, B cell receptor, and J. Exp. Med. 1995; PubMed Scopus Google Scholar). We that during the of platelet aggregation, PECAM-1 with receptors such as studies have that of the SH2 domains of a can to the direct of its S. J. Biol. Chem. 1994; 269: Full Text PDF PubMed Google Scholar). study the of the SH2 domains and induction of activation of we the of PECAM-1 peptides the tyrosine residues within the cytoplasmic domain of PECAM-1 to activation of recombinant SHP-1. studies demonstrate SHP-2, two tyrosine-phosphorylated and peptides to SHP-1 catalytic higher of SHP-1 activation with tyrosine-phosphorylated peptide is with the by the of the 2 of the tyrosine in the SHP-1 binding motif D.N. W. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). studies have the structural of association and for phosphatase the of further is that SHP-1 have a different role with SHP-2 to the PECAM-1 These SHP-2, and have to either or on cellular signaling SHP-1 as having a role in signaling by receptor Cell. Biol. PubMed Google U. S.J. J. Exp. Med. 1996; PubMed Scopus Google Scholar, S. A. 1996; PubMed Scopus Google receptor S. Cell. Biol. 1996; PubMed Scopus Google B cell receptor 1995; Full Text PDF PubMed Scopus Google and cell receptor I. J. Exp. Med. 1996; PubMed Scopus Google Scholar). to be by SHP-1 in activation of the protein kinase L. D. J. Biol. Chem. 1996; 271: Full Text Full Text PDF PubMed Scopus Google in and of M. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google and in and activation of the protein-tyrosine kinases such as pp60c-src in hematopoietic cells J.S. D.R. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). a of both SHP-1 and SHP-2 for the binding in PECAM-1 at in a hematopoietic cell it is that the be by its binding its induction of its tyrosine phosphorylation, and its to effects on signaling be by in its in to extracellular and of a of tyrosine-phosphorylated SHP-1 to the cytoskeletal is within of integrin αIIbβ3-mediated platelet aggregation A. J.M. 1994; PubMed Scopus Google A. J.M. J. 1995; PubMed Scopus Google Scholar). In we that the of SHP-1 to the during platelet aggregation with kinetics to with PECAM-1 and pp60c-src This in cellular of SHP-1 the effects of SHP-2, which is into the or with the during platelet aggregation Y. M. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar). within the of the SHP-1 be the for In to its SHP-1 tyrosine-phosphorylated during platelet aggregation as predominantly in the and to a in the A. J.M. 1994; PubMed Scopus Google A. J.M. J. 1995; PubMed Scopus Google Scholar). results suggest that tyrosine-phosphorylated SHP-1 and different downstream signaling molecules than SHP-2, PECAM-1 with cell signaling downstream of an SHP-1 SH2 domain interaction is the studies have that in hematopoietic SHP-1 can associate with a that is with such as the with extracellular kinase activation M. J. Biol. Chem. 1996; 271: Full Text Full Text PDF PubMed Scopus Google Scholar). interaction of PECAM-1 with tyrosine-phosphorylated SHP-1 a to the protein kinase in that PECAM-1 and SHP-1 association extracellular kinase phosphorylation and which occurs within 2 of integrin αIIbβ3-mediated platelet aggregation, is with to this downstream signaling In we have that in to SHP-2, can physically associate via a common motif phosphorylation of SHP-1 and SHP-2 to be to its binding to since the PECAM-1 phosphopeptides encompassing tyrosine residues and can as for the catalytic activation of the of SHP-1 and SHP-2 with PECAM-1 represents a the which or inhibitory signals to cellular signaling We to Newman for and the of and to for the SHP-1 and SHP-2 in