Subscribe to RSS
DOI: 10.1055/s-0037-1614177
Structure and Function of Murine αIIbβ3 (GPIIb/IIIa): Studies Using Monoclonal Antibodies and β3-null Mice
This study was supported in part by grants 19278 and 54469 from the National Heart, Lung and Blood Institute, and in part by a grant from the Swiss National Science Foundation (NF-3100.053408.98).Publication History
Received
16 May 2000
Accepted after revision
11 July 2000
Publication Date:
13 December 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/200006/lookinside/thumbnails/10-1055-s-0037-1614177-1.jpg)
Summary
The αIIbβ3 receptor (GPIIb/IIIa) is the only platelet-specific integrin receptor and the most abundant adhesion/aggregation receptor on the surface of human platelets. Since mice are increasingly being used as models of human disease, we analyzed the structure and function of murine platelet αIIbβ3, utilizing both β3 integrin-deficient mice, who have a phenotype that resembles Glanzmann thrombasthenia, and our hamster monoclonal antibody (mAb) 1B5 to murine αIIbβ3. By immunoblot analysis, flow cytometry, and mAb binding studies, mouse platelets express abundant amounts of αIIbβ3 (60-80,000 copies/platelet). Like their human counterparts, murine αIIb and β3 exhibit different electrophoretic motilities under nonreducing (αIIb 135k Da; β3 92k Da) and reducing (αIIb 120k Da; β3 108k Da) conditions, and the αIIbβ3 complex is dissociated by EDTA at pH 8 and 37 ºC. Murine β3 is less susceptible to proteolysis by plasmin than is human β3. In addition to defective platelet aggregation, mouse platelets lacking αIIbβ3 and αVβ3 are unable to adhere to fibrinogen and prothrombin, but retain the ability to adhere to fibronectin and collagen. Following platelet activation, β3-null platelets express slightly less P-selectin than do wild-type mouse platelets. Moreover, β3-null platelets have altered tyrosine phosphorylation patterns following thrombinand collagen-induced aggregation. These results suggest fundamental similarities between human and mouse platelet activation and aggregation, but delineate subtle differences that need to be considered when comparing studies from mice and humans.
* DAT and SSS contributed equally to this work
-
References
- 1 Du X, Ginsberg MH. Integrin αIIbβ3 and platelet function. Thromb Haemost 1997; 78: 96-100.
- 2 Plow EF, Ginsberg MH. Cellular adhesion: GPIIb-IIIa as a prototypic adhesion receptor. Prog Hemost Thromb 1989; 09: 117-56.
- 3 Coller BS, Seligsohn U, Peretz H, Newman PJ. Glanzmann thrombasthenia: new insights from an historical perspective. Semin Hematol 1994; 31: 301-11.
- 4 George JN, Caen JP, Nurden AT. Glanzmann’s thrombasthenia: the spectrum of clinical disease. Blood 1990; 75: 1383-95.
- 5 Shattil SJ, Kashiwagi H, Pampori N. Integrin signaling: the platelet paradigm. Blood 1998; 91: 2645-57.
- 6 Majzoub JA, Muglia LJ. Knockout mice. N Engl J Med 1996; 334: 904-7.
- 7 Hynes RO, Bader BL. Targeted mutations in integrins and their ligands: their implications for vascular biology. Thromb Haemost 1997; 78: 83-7.
- 8 Hodivala-Dilke KM, McHugh K, Tsakiris DA, Rayburn H, Crowley D, Ullman-Culliere M, Ross FP, Coller BS, Teitelbaum S, Hynes RO. β3 integrin deficient mice: a model for Glanzmann thrombasthenia showing placental defects and reduced survival. J Clin Invest 1999; 103: 229-38.
- 9 Thornton MA, Poncz M. Characterization of the murine platelet alphaIIβ gene and encoded cDNA. Blood 1999; 94: 3947-50.
- 10 Cieutat AM, Rosa JP, Letourneur F, Poncz M, Rifat S. A comparative analysis of cDNA-derived sequences for rat and mouse beta 3 integrins (GPIIIa) with their human counterpart. Biochem Biophys Res Commun 1993; 193: 771-8.
- 11 Lengweiler S, Smyth SS, Jirouskova M, Scudder L, Park H, Moran T, Coller BS. Preparation of monoclonal antibodies to murine platelets from hamster-mouse interspecies hybridomas. Biochem Biophys Res Commun 1999; 262: 167-73.
- 12 Coller BS, Seligsohn U, Little PA. Type I Glanzmann thrombasthenia patients from the Iraqi-Jewish and Arab populations in Israel can be differentiated by platelet glycoprotein IIIa immunoblot analysis. Blood 1987; 69: 1696-703.
- 13 Yasuda M, Hasunuma Y, Adachi H, Sekine C, Sakanishi T, Hashimoto H, Ra C, Yagita H, Okumura K. Expression and function of fibronectin binding integrins on rat mast cells. Int Immunol 1995; 07: 251-8.
- 14 Schultz JF, Armant DR. Beta 1- and beta 3-class integrins mediate fibronectin binding activity at the surface of developing mouse periimplantation blastocysts Regulation by ligand-induced mobilization of stored receptor. J Biol Chem 1995; 270: 11522-31.
- 15 Frenette PS, Johnson RC, Hynes RO, Wagner DD. Platelets roll on stimulated endothelium in vivo: an interaction mediated by endothelial P-selectin. Proc Natl Acad Sci USA 1995; 92: 7450-74.
- 16 Beer J, Coller BS. Evidence that platelet glycoprotein IIIa has a large disulfide bonded loop that is susceptible to proteolytic cleavage. J Biol Chem 1989; 264: 17564-73.
- 17 Scudder LE, Kalomiris EL, Coller BS. Preparation and functional characterization of monoclonal antibodies against glycoprotein Ib. Methods Enzymol 1992; 215: 295-311.
- 18 Phillips DR, Agin PP. Platelet plasma membrane glycoproteins Evidence for the presence of nonequivalent disulfide bonds using nonreduced-reduced two-dimensional gel electrophoresis. J Biol Chem 1977; 252: 2121-6.
- 19 Clark EA, Shattil SJ, Ginsberg MH, Bolen J, Brugge JS. Regulation of the protein tyrosine kinase p72syk by platelet agonists and the integrin alpha IIb beta 3. J Biol Chem 1994; 269: 28859-64.
- 20 Tsakiris DA, Scudder L, Hodivala-Dilke K, Hynes RO, Coller BS. Hemostasis in the mouse (Mus musculus): a review. Thromb Haemost 1999; 81: 177-88.
- 21 Gachet C, Stierle A, Ohlmann P, Lanza F, Hanau D, Cazenave JP. Normal ADP-induced aggregation and absence of dissociation of the membrane GP IIb-IIIa complex of intact rat platelets pretreated with EDTA. Thromb Haemost 1991; 66: 246-53.
- 22 Wagner CL, Mascelli MA, Neblock DS, Weisman HF, Coller BS, Jordan RE. Analysis of GPIIb/IIIa receptor number by quantification of 7E3 binding to human platelets. Blood 1996; 88: 907-14.