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DOI: 10.1055/s-0037-1614429
Clustering of Integrin αIIb-β3Differently Regulates Tyrosine Phosphorylation of pp72syk, PLCγ2 and pp125FAKin Concanavalin A-stimulated Platelets
Correspondence to:
Publication History
Received29 April 1998
Accepted after revision22 September 1998
Publication Date:
08 December 2017 (online)
Summary
Tyrosine phosphorylation of the non-receptor tyrosine kinases pp72sykand pp125FAKand of the γ2 isoform of phospholipase C (PLCγ2) in human platelets stimulated with the lectin Concanavalin A was investigated. Concanavalin A induced the rapid tyrosine phosphorylation of pp72sykand PLCγ2 with a similar kinetics, while tyrosine phosphorylation of pp125FAKoccurred in a later phase of platelet activation. When compared with other platelet agonists, Concanavalin A revealed to be at least as potent as collagen in inducing tyrosine phosphorylation of PLCγ2 and pp125FAK, while tyrosine phosphorylation of pp72sykinduced by the lectin was much stronger than that induced by thrombin or collagen. Concanavalin A-induced tyrosine phosphorylation of pp72syk, PLCγ2 and pp125FAKwas not dependent on platelet aggregation as it occurred normally even in the absence of sample stirring and when fibrinogen binding to integrin αIIb-β3was inhibited by the peptide RGDS. Tyrosine phosphorylation of pp72syk, PLCγ2 and pp125FAKrequired the binding of the lectin to the platelet surface, but was not observed in platelets treated with succinyl-Concanavalin A, a derivative of the lectin that interacts with the same receptors but does not promote clustering of membrane glycoproteins. Moreover, the aggregation-independent tyrosine phosphorylation of pp125FAKand pp72sykinduced by Concanavalin A required the expression of integrin αIIb-β3on the platelet surface as it was strongly inhibited in platelets from patients affected by Glanzmann thrombasthenia. By contrast, tyrosine phosphorylation of PLCγ2 occurred normally also in thrombasthenic platelets stimulated with Concanavalin A. These results demonstrate that, even in the absence of aggregation, the clustering of integrin αIIb-β3induced by Concanavalin A on the platelet surface directly promotes tyrosine phosphorylation of pp72sykand pp125FAKand provide further evidence that the oligomerization of the fibrinogen receptor promoted by its natural ligand during platelet aggregation may be responsible for the tyrosine phosphorylation of these proteins induced by physiological agonists.
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References
- 1 Jackson SP, Schoenwaelder SM, Yuan Y, Salem HH, Cooray P. Non-receptor protein tyrosine kinases and phosphatases in human platelets. Thromb Haemost 1996; 76: 640-50.
- 2 Torti M, Lapetina EG. Role of rap1B and the p21ras GTPase activating protein in the regulation of phospholipase Cγ1 in human platelets. Proc Natl Acad Sci USA 1992; 89: 7796-800.
- 3 Vostal JG, Shulman NR. Vinculin is a major platelet protein that undergoes Ca2+-dependent tyrosine phosphorylation. Biochem J 1993; 294: 675-80.
- 4 Taniguchi T, Kitagawa H, Yasue S, Yanagi S, Sakai K, Asahi M, Otha S, Takeuchi F, Nakamura S, Yamamura H. Protein-tyrosine kinase p72syk is activated by thrombin and is negatively regulated through Ca2+ mobilization in platelets. J Biol Chem 1993; 268: 2277-9.
- 5 Clark EA, Shattil SJ, Ginsberg MH, Bolen J, Brugge JS. Regulation of the protein tyrosine kinase pp72syk by platelet agonists and the integrin αIIb-β3 . J Biol Chem 1994; 269: 28859-64.
- 6 Li RY, Gaits F, Ragab A, Ragab-Thomas JMF, Chap H. Tyrosine phosphorylation of an SH2-containing protein tyrosine phosphatase is coupled to platelet thrombin receptor via a pertussis toxin-sensitive heterotrimeric G-protein. EMBO J 1995; 14: 2519-26.
- 7 Ferrell Jr JE, Martin GS. Tyrosine-specific protein phosphorylation is regulated by glycoprotein IIb-IIIa in platelets. Proc Natl Acad Sci USA 1989; 86: 2234-8.
- 8 Golden A, Brugge JS, Shattil SJ. Role of platelet membrane glycoprotein IIb-IIIa in agonist-induced tyrosine phosphorylation of platelet proteins. J Cell Biol 1990; 111: 3117-27.
- 9 Lipfert L, Haimovich B, Schaller MB, Cobb BS, Parsons JT, Brugge JS. Integrin-dependent phosphorylation and activation of the protein tyrosine kinase pp125FAK in platelets. J Cell Biol 1992; 119: 905-12.
- 10 Haimovich B, Lipfert L, Brugge JS, Shattil SJ. Tyrosine phosphorylation and cytoskeletal reorganisation in platelets are triggered by interaction of integrin receptors with their immobilised ligands. J Biol Chem 1993; 268: 15868-77.
- 11 Huang M, Lipfert L, Cunningham M, Brugge JS, Ginsberg MH, Shattil SJ. Adhesive ligand binding to integrin αIIb-β3 stimulates tyrosine phosphorylation of novel protein substrates before phosphorylation of pp125FAK . J Cell Biol 1993; 122: 473-83.
- 12 Gao J, Zoller KE, Ginsberg MH, Brugge JS, Shattil SJ. Regulation of the pp72syk protein tyrosine kinase by platelet integrin αIIb-β3 . EMBO J 1997; 16: 6414-25.
- 13 Shattil SJ, Haimovich B, Cunningham M, Lipfert L, Parsons JT, Ginsberg MH, Brugge JS. Tyrosine phosphorylation of pp125FAK in platelets requires coordinated signaling through integrin and agonist receptors. J Biol Chem 1994; 269: 14738-45.
- 14 Isenberg WM, McEver RP, Phillips DR, Schuman MA, Bainton DF. The platelet fibrinogen receptor: an immunogold-surface replica study of agonist-induced ligand binding and receptor clustering. J Cell Biol 1987; 104: 1655-63.
- 15 Lawler J. The structural and functional properties of thrombospondin. Blood 1986; 67: 1197-209.
- 16 Phillips DR, Jennings LK, Edwards HH. Identification of membrane proteins mediating the interaction of human platelets. J Cell Biol 1980; 86: 77-86.
- 17 Newman PJ, Hillery CA, Albrecht R, Parise LV, Berndt MC, Mazurov AV, Dunlop L C, Zhang J, Rittenhouse SE. Activation-dependent changes in human platelet PECAM-1: phosphorylation, cytoskeletal association, and surface membrane redistribution. J Cell Biol 1992; 119: 239-46.
- 18 Torti M, Balduini C, Ramaschi G, Sinigaglia F. Stimulation of human platelets with Concanavalin A involves phospholipase C activation. Cell Biochem Funct 1992; 10: 53-9.
- 19 Wheeler ME, Gerrard JM, Carrol RC. Reciprocal transmembranous receptor-cytoskeleton interaction in Concanavalin A-activated platelets. J Cell Biol 1985; 101: 993-1000.
- 20 Fitzgerald LA, Leung B, Phillips DR. A method for purifying platelet membrane glycoprotein IIb-IIIa complex. Anal Biochem 1985; 151: 169-77.
- 21 Kakaiya RM, Kiroly TL, Cable RG. Concanavalin A induces patching/ capping of the platelet membrane glycoprotein IIb-IIIa complex. Thromb Haemost 1988; 59: 281-3.
- 22 Torti M, Ramaschi G, Sinigaglia F, Balduini C. Dual mechanism of protein tyrosine phosphorylation in Concanavalin A-stimulated platelets. J Cell Biochem 1995; 57: 30-8.
- 23 Keeley PJ, Parise LV. The α2β1 integrin is a necessary co-receptor for collagen-induced activation of Syk and the subsequent phosphorylation of phospholipase Cγ2 in platelets. J Biol Chem 1996; 272: 26668-76.
- 24 Tate BF, Rittenhouse SE. Thrombin activation of human platelets causes tyrosine phosphorylation of PLC-γ2. Biochim Biophys Acta 1993; 1178: 281-5.
- 25 Daniel JL, Dangelmaier C, Smith JB. Evidence for a role for tyrosine phosphorylation of phospholipase Cγ2 in collagen-induced platelet cytosolic calcium mobilization. Biochem J 1994; 302: 617-22.
- 26 Moroi M, Jung SM. Selective staining of human platelet glycoproteins using nitrocellulose transfer of electrophoresed proteins and peroxidase-conjugated lectins. Biochim Biophys Acta 1984; 798: 295-301.
- 27 Gunther GR, Wang JL, Yahara I, Cunningham BA, Edelman GM. Concanavalin A derivatives with altered biological activities. Proc Natl Acad Sci USA 1973; 70: 1012-6.
- 28 Rhee SG. Inositol phospholipid-specific phospholipase C: interaction of the γ1 isoform with tyrosine kinase. Trends Biochem Sci 1991; 16: 297-301.
- 29 Asselin J, Gibbins JM, Achison M, Lee YH, Morton LF, Farndale RW, Barnes MJ, Watson SP. A collagen-like peptide stimulates tyrosine phosphorylation of syk and phospholipase Cγ2 in platelets independent of integrin α2β1 . Blood 1997; 89: 1235-42.
- 30 Torti M, Ramaschi G, Montsarrat N, Sinigaglia F, Balduini C, Plantavid M, Breton M, Chap H, Mauco G. Evidence for a glycoprotein IIb-IIIa- and aggregation-independent mechanism of phosphatidylinositol 3’,4’-bisphosphate synthesis in human platelets. J Biol Chem 1995; 270: 13179-85.
- 31 Indig FE, Diaz-Gonzales F, Ginsberg MH. Analysis of tetraspanin CD9-in-tegrin αIIb-β3 (GP IIb-IIIa) complex in platelet membranes and transfected cells. Biochem J 1997; 327: 291-8.
- 32 Chacko GW, Duchemin A, Coggeshall KM, Osborne JM, Brandt JT, Anderson CL. Clustering of the platelet Fcγ receptor induces noncovalent association with the tyrosine kinase p72syk . J Biol Chem 1994; 269: 32435-40.
- 33 Ichinohe T, Takayama H, Yasuharu E, Arai M, Yamamoto N, Takahashi H, Okuma M. Collagen-stimulated activation of Syk but not c-Src is severely compromised in human platelets lacking membrane glycoprotein VI. J Biol Chem 1997; 272: 63-8.
Correspondence to:
-
References
- 1 Jackson SP, Schoenwaelder SM, Yuan Y, Salem HH, Cooray P. Non-receptor protein tyrosine kinases and phosphatases in human platelets. Thromb Haemost 1996; 76: 640-50.
- 2 Torti M, Lapetina EG. Role of rap1B and the p21ras GTPase activating protein in the regulation of phospholipase Cγ1 in human platelets. Proc Natl Acad Sci USA 1992; 89: 7796-800.
- 3 Vostal JG, Shulman NR. Vinculin is a major platelet protein that undergoes Ca2+-dependent tyrosine phosphorylation. Biochem J 1993; 294: 675-80.
- 4 Taniguchi T, Kitagawa H, Yasue S, Yanagi S, Sakai K, Asahi M, Otha S, Takeuchi F, Nakamura S, Yamamura H. Protein-tyrosine kinase p72syk is activated by thrombin and is negatively regulated through Ca2+ mobilization in platelets. J Biol Chem 1993; 268: 2277-9.
- 5 Clark EA, Shattil SJ, Ginsberg MH, Bolen J, Brugge JS. Regulation of the protein tyrosine kinase pp72syk by platelet agonists and the integrin αIIb-β3 . J Biol Chem 1994; 269: 28859-64.
- 6 Li RY, Gaits F, Ragab A, Ragab-Thomas JMF, Chap H. Tyrosine phosphorylation of an SH2-containing protein tyrosine phosphatase is coupled to platelet thrombin receptor via a pertussis toxin-sensitive heterotrimeric G-protein. EMBO J 1995; 14: 2519-26.
- 7 Ferrell Jr JE, Martin GS. Tyrosine-specific protein phosphorylation is regulated by glycoprotein IIb-IIIa in platelets. Proc Natl Acad Sci USA 1989; 86: 2234-8.
- 8 Golden A, Brugge JS, Shattil SJ. Role of platelet membrane glycoprotein IIb-IIIa in agonist-induced tyrosine phosphorylation of platelet proteins. J Cell Biol 1990; 111: 3117-27.
- 9 Lipfert L, Haimovich B, Schaller MB, Cobb BS, Parsons JT, Brugge JS. Integrin-dependent phosphorylation and activation of the protein tyrosine kinase pp125FAK in platelets. J Cell Biol 1992; 119: 905-12.
- 10 Haimovich B, Lipfert L, Brugge JS, Shattil SJ. Tyrosine phosphorylation and cytoskeletal reorganisation in platelets are triggered by interaction of integrin receptors with their immobilised ligands. J Biol Chem 1993; 268: 15868-77.
- 11 Huang M, Lipfert L, Cunningham M, Brugge JS, Ginsberg MH, Shattil SJ. Adhesive ligand binding to integrin αIIb-β3 stimulates tyrosine phosphorylation of novel protein substrates before phosphorylation of pp125FAK . J Cell Biol 1993; 122: 473-83.
- 12 Gao J, Zoller KE, Ginsberg MH, Brugge JS, Shattil SJ. Regulation of the pp72syk protein tyrosine kinase by platelet integrin αIIb-β3 . EMBO J 1997; 16: 6414-25.
- 13 Shattil SJ, Haimovich B, Cunningham M, Lipfert L, Parsons JT, Ginsberg MH, Brugge JS. Tyrosine phosphorylation of pp125FAK in platelets requires coordinated signaling through integrin and agonist receptors. J Biol Chem 1994; 269: 14738-45.
- 14 Isenberg WM, McEver RP, Phillips DR, Schuman MA, Bainton DF. The platelet fibrinogen receptor: an immunogold-surface replica study of agonist-induced ligand binding and receptor clustering. J Cell Biol 1987; 104: 1655-63.
- 15 Lawler J. The structural and functional properties of thrombospondin. Blood 1986; 67: 1197-209.
- 16 Phillips DR, Jennings LK, Edwards HH. Identification of membrane proteins mediating the interaction of human platelets. J Cell Biol 1980; 86: 77-86.
- 17 Newman PJ, Hillery CA, Albrecht R, Parise LV, Berndt MC, Mazurov AV, Dunlop L C, Zhang J, Rittenhouse SE. Activation-dependent changes in human platelet PECAM-1: phosphorylation, cytoskeletal association, and surface membrane redistribution. J Cell Biol 1992; 119: 239-46.
- 18 Torti M, Balduini C, Ramaschi G, Sinigaglia F. Stimulation of human platelets with Concanavalin A involves phospholipase C activation. Cell Biochem Funct 1992; 10: 53-9.
- 19 Wheeler ME, Gerrard JM, Carrol RC. Reciprocal transmembranous receptor-cytoskeleton interaction in Concanavalin A-activated platelets. J Cell Biol 1985; 101: 993-1000.
- 20 Fitzgerald LA, Leung B, Phillips DR. A method for purifying platelet membrane glycoprotein IIb-IIIa complex. Anal Biochem 1985; 151: 169-77.
- 21 Kakaiya RM, Kiroly TL, Cable RG. Concanavalin A induces patching/ capping of the platelet membrane glycoprotein IIb-IIIa complex. Thromb Haemost 1988; 59: 281-3.
- 22 Torti M, Ramaschi G, Sinigaglia F, Balduini C. Dual mechanism of protein tyrosine phosphorylation in Concanavalin A-stimulated platelets. J Cell Biochem 1995; 57: 30-8.
- 23 Keeley PJ, Parise LV. The α2β1 integrin is a necessary co-receptor for collagen-induced activation of Syk and the subsequent phosphorylation of phospholipase Cγ2 in platelets. J Biol Chem 1996; 272: 26668-76.
- 24 Tate BF, Rittenhouse SE. Thrombin activation of human platelets causes tyrosine phosphorylation of PLC-γ2. Biochim Biophys Acta 1993; 1178: 281-5.
- 25 Daniel JL, Dangelmaier C, Smith JB. Evidence for a role for tyrosine phosphorylation of phospholipase Cγ2 in collagen-induced platelet cytosolic calcium mobilization. Biochem J 1994; 302: 617-22.
- 26 Moroi M, Jung SM. Selective staining of human platelet glycoproteins using nitrocellulose transfer of electrophoresed proteins and peroxidase-conjugated lectins. Biochim Biophys Acta 1984; 798: 295-301.
- 27 Gunther GR, Wang JL, Yahara I, Cunningham BA, Edelman GM. Concanavalin A derivatives with altered biological activities. Proc Natl Acad Sci USA 1973; 70: 1012-6.
- 28 Rhee SG. Inositol phospholipid-specific phospholipase C: interaction of the γ1 isoform with tyrosine kinase. Trends Biochem Sci 1991; 16: 297-301.
- 29 Asselin J, Gibbins JM, Achison M, Lee YH, Morton LF, Farndale RW, Barnes MJ, Watson SP. A collagen-like peptide stimulates tyrosine phosphorylation of syk and phospholipase Cγ2 in platelets independent of integrin α2β1 . Blood 1997; 89: 1235-42.
- 30 Torti M, Ramaschi G, Montsarrat N, Sinigaglia F, Balduini C, Plantavid M, Breton M, Chap H, Mauco G. Evidence for a glycoprotein IIb-IIIa- and aggregation-independent mechanism of phosphatidylinositol 3’,4’-bisphosphate synthesis in human platelets. J Biol Chem 1995; 270: 13179-85.
- 31 Indig FE, Diaz-Gonzales F, Ginsberg MH. Analysis of tetraspanin CD9-in-tegrin αIIb-β3 (GP IIb-IIIa) complex in platelet membranes and transfected cells. Biochem J 1997; 327: 291-8.
- 32 Chacko GW, Duchemin A, Coggeshall KM, Osborne JM, Brandt JT, Anderson CL. Clustering of the platelet Fcγ receptor induces noncovalent association with the tyrosine kinase p72syk . J Biol Chem 1994; 269: 32435-40.
- 33 Ichinohe T, Takayama H, Yasuharu E, Arai M, Yamamoto N, Takahashi H, Okuma M. Collagen-stimulated activation of Syk but not c-Src is severely compromised in human platelets lacking membrane glycoprotein VI. J Biol Chem 1997; 272: 63-8.