The Cleaved Peptide of PAR1 Results in a Redistribution of the Platelet Surface GPIb-IX-V Complex to the Surface-Connected Canalicular System

Mark I. Furman; Paquita Nurden; Michael C. Berndt; Alan T. Nurden; Stephen E. Benoit; Marc R. Barnard; Frederick A. Ofosu; Alan D. Michelson

doi:10.1055/s-0037-1614134

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2000; 84(05): 897-903
DOI: 10.1055/s-0037-1614134

Review Article

Schattauer GmbH

The Cleaved Peptide of PAR1 Results in a Redistribution of the Platelet Surface GPIb-IX-V Complex to the Surface-Connected Canalicular System

Mark I. Furman

¹From the Center for Platelet Function Studies, Division of Cardiovascular Medicine, Department of Medicine, UMass Memorial Health Care, University of Massachusetts Medical School, Worcester, Massachusetts, USA

,

Paquita Nurden

²UMR 5533 CNRS, Hopital Cardiologique, Pessac, France

,

Michael C. Berndt

³Baker Medical Research Institute, Prahran, Australia

,

Alan T. Nurden

²UMR 5533 CNRS, Hopital Cardiologique, Pessac, France

,

Stephen E. Benoit

¹From the Center for Platelet Function Studies, Division of Cardiovascular Medicine, Department of Medicine, UMass Memorial Health Care, University of Massachusetts Medical School, Worcester, Massachusetts, USA

⁴Center for Platelet Function Studies, Department of Pediatrics, UMass Memorial Health Care, University of Massachusetts Medical School, orcester, Massachusetts, USA

,

Marc R. Barnard

⁴Center for Platelet Function Studies, Department of Pediatrics, UMass Memorial Health Care, University of Massachusetts Medical School, orcester, Massachusetts, USA

,

Frederick A. Ofosu

⁵Department of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada

,

Alan D. Michelson

⁴Center for Platelet Function Studies, Department of Pediatrics, UMass Memorial Health Care, University of Massachusetts Medical School, orcester, Massachusetts, USA

› Author Affiliations

Abstract

Summary

The only known function of the 41 amino acid cleaved peptide (TR_1-41) of the seven transmembrane domain thrombin receptor (PAR1) is to activate platelets (as determined by aggregation, surface P-selectin, and fibrinogen binding to activated GPIIb-IIIa). We now demonstrate that TR_1-41 results in a concentration-dependent decrease in the platelet surface expression of each component of the GPIb-IX-V complex, as determined by flow cytometry with a panel of monoclonal antibodies (including 6D1, directed against the von Willebrand factor binding site on GPIbα, and TM60, directed against the thrombin binding site on GPIbα). TR_1-41 also decreased ristocetin-induced platelet agglutination. Immunoblotting after incubation of platelets with TR_1-41 revealed neither a loss of platelet GPIb nor increase in supernatant GPIb fragments. As demonstrated by immunoelectron microscopy, TR_1-41 resulted in a redistribution of GPIb, GPIX, and GPV from the platelet surface to the surface-connected canalicular system (SCCS). In summary, the cleaved peptide (TR_1-41) of PAR1 results in a redistribution of the platelet surface GPIb-IX-V complex to the SCCS, thereby negatively regulating the GPIbα binding sites for von Willebrand factor and thrombin.

Key words

Thrombin - thrombin receptor - von Willebrand factor - peptide - flow cytometry

Full Text

References

References
1 Ruggeri ZM. The platelet glycoprotein Ib-IX complex. Prog Hemost Thromb 1991; 10: 35-68.
2 Roth GJ. Developing relationships: arterial platelet adhesion, glycoprotein Ib, and leucine-rich glycoproteins. Blood 1991; 77: 5-19.
3 Lopez JA. The platelet glycoprotein Ib-IX complex. Blood Coagul.Fibrinolysis 1994; 05: 97-119.
4 Michelson AD. CD42 cluster workshop report. In: Schlossman SF, Boumsell L, Gilks W. et al, eds. Leucocyte typing V. Oxford: Oxford University Press; 1995: 1309-10.
5 Wenger RH, Wicki AN, Kieffer N, Adolph S, Hameister H, Clemetson KJ. The 5’ flanking region and chromosomal localization of the gene encoding human platelet membrane glycoprotein Ib alpha. Gene 1989; 85: 517-24.
6 Hickey MJ, Deaven LL, Roth GJ. Human platelet glycoprotein IX. Characterization of cDNA and localization of the gene to chromosome 3. FEBS Lett 1990; 274: 189-92.
7 Modderman PW, Admiraal LG, Sonnenberg A, von dem Borne AE. Glycoproteins V and Ib-IX form a noncovalent complex in the platelet membrane. J Biol Chem 1992; 267: 364-9.
8 Coller BS, Peerschke EI, Scudder LE, Sullivan CA. Studies with a murine monoclonal antibody that abolishes ristocetin-induced binding of von Willebrand factor to platelets: additional evidence in support of GPIb as a platelet receptor for von Willebrand factor. Blood 1983; 61: 99-110.
9 Du X, Beutler L, Ruan C, Castaldi PA, Berndt MC. Glycoprotein Ib and glycoprotein IX are fully complexed in the intact platelet membrane. Blood 1987; 69: 1524-7.
10 Romo GM, Dong JF, Schade AJ, Gardiner EE, Kansas GS, Li CQ, McIntire LV, Berndt MC, Lopez JA. The glycoprotein Ib-IX-V complex is a platelet counterreceptor for P-selectin. J Exp Med 1999; 190 (06) 803-14.
11 Harmon JT, Jamieson GA. The glycocalicin portion of platelet glycoprotein Ib expresses both high and moderate affinity receptor sites for thrombin. J Biol Chem 1986; 261: 13224-9.
12 Yamamoto N, Greco NJ, Barnard MR. et al. Glycoprotein Ib (GPIb)-dependent and GPIb-independent pathways of thrombin-induced platelet activation. Blood 1991; 77: 1740-8.
13 Gralnick HR, Williams S, McKeown LP, Hansmann K, Fenton JW, Krutzsch H. High-affinity alpha-thrombin binding to platelet glycoprotein Ib alpha: identification of two binding domains. Proc Natl Acad Sci USA 1994; 91: 6334-8.
14 De Marco L, Mazzucato M, Masotti A, Ruggeri ZM. Localization and characterization of an alpha-thrombin-binding site on platelet glycoprotein Ib alpha. J Biol Chem 1994; 269: 6478-84.
15 Fox JEB, Berndt MC. Cyclic AMP-dependent phosphorylation of glycoprotein Ib inhibits collagen-induced polymerization of actin in platelets. J Biol Chem 1989; 264: 9520-6.
16 Dong JF, Sae-Tung G, Lopez JA. Role of glycoprotein V in the formation of the platelet high- affinity thrombin-binding site. Blood 1997; 89: 4355-63.
17 Hansen SR, Harker LA. Interruption of acute platelet-dependent thrombosis by the synthetic antithrombin PPACK. Proc Natl Acad Sci USA 1988; 85: 3184-8.
18 Eidt JF, Allison P, Nobel S. et al. Thrombin is an important mediator of platelet aggregation in stenosed canine coronary arteries with endothelial injury. J Clin Inves 1989; 84: 18-27.
19 Kelly AB, Marzec UM, Krupski W. et al. Hirudin interruption of heparinresistant arterial thrombus formation in baboons. Blood 1991; 77: 1006-12.
20 Vu T-KH, Hung DT, Wheaton VI, Coughlin SR. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 1991; 64: 1057-68.
21 Furman MI, Liu L, Benoit SE, Becker RC, Barnard MR, Michelson AD. The cleaved peptide of the thrombin receptor is a strong platelet agonist. Proc Natl Acad Sci USA 1998; 95: 3082-7.
22 Michelson AD, Loscalzo J, Melnick B, Coller BS, Handin RI. Partial characterization of a binding site for von Willebrand factor on glycocalicin. Blood 1986; 67: 19-26.
23 Yamamoto N, Kitagawa H, Tanoue K, Yamazaki H. Monoclonal antibody to glycoprotein Ib inhibits both thrombin- and ristocetin-induced platelet aggregations. Thromb Res 1985; 39: 751-9.
24 Yamamoto K, Yamamoto N, Kitagawa H, Tanoue K, Kosaki G, Yamazaki H. Localization of a thrombin-binding site on human platelet membrane glycoprotein Ib determined by a monoclonal antibody. Thromb Haemost 1986; 55: 162-7.
25 Katagiri Y, Hayashi Y, Yamamoto K, Tanoue K, Kosaki G, Yamazaki H. Localization of von Willebrand factor and thrombin-interactive domains on human platelet glycoprotein Ib. Thromb Haemost 1990; 63: 122-6.
26 Michelson AD, Barnard MR. Plasmin-induced redistribution of platelet glycoprotein Ib. Blood 1990; 76: 2005-10.
27 Michelson AD. Flow cytometry: a clinical test of platelet function. Blood 1996; 87: 4925-36.
28 Berndt MC, Phillips DR. Purification and preliminary physicochemical characterization of human platelet membrane glycoprotein V. J Biol Chem 1981; 256: 59-65.
29 Ward CM, Andrews RK, Smith AI, Berndt MC. Mocarhagin, a novel cobra venom metalloproteinase, cleaves the platelet von Willebrand factor receptor glycoprotein Ibα. Identification of the sulfated tyrosine/anionic sequence Tyr-276-Glu-282 of glycoprotein Ibα as a binding site for von Willebrand factor and α-thrombin. Biochemistry 1996; 35: 4929-38.
30 Berndt MC, Gregory C, Kabral A, Zola H, Fournier D, Castaldi PA. Purification and preliminary characterization of the glycoprotein Ib complex in the human platelet membrane. Eur J Biochem 1985; 151: 637-49.
31 Hourdille P, Heilmann E, Combrie R, Winckler J, Clemetson KJ, Nurden AT. Thrombin induces a rapid redistribution of glycoprotein Ib-IX complexes within the membrane systems of activated human platelets. Blood 1990; 76: 1503-13.
32 Michelson AD, Benoit SE, Furman MI, Barnard MR, Nurden P, Nurden AT. The platelet surface expression of glycoprotein V is regulated by two independent mechanisms: Proteolysis and a reversible cytoskeletal-mediated redistribution to the surface-connected canalicular system. Blood 1996; 87: 1396-408.
33 Poujol C, Nurden AT, Nurden P. Ultrastructural analysis of the distribution of the vitronectin receptor (alpha v beta 3) in human platelets and megakaryocytes reveals an intracellular pool and labelling of the alpha-granule membrane. Br J Haematol 1997; 96: 823-35.
34 Nurden A, Cazes E, Bihour C. et al. Confirmation that GPIb-IX complexes have a reduced surface distribution on platelets activated by thrombin and TRAP-14-mer peptide. Br J Haematol 1995; 90: 645-54.
35 Liu L, Freedman J, Hornstein A, Fenton JW, Song Y, Ofosu FA. Binding of thrombin to the G-protein-linked receptor, and not to glycoprotein Ib, precedes thrombin-mediated platelet activation. J Biol Chem 1997; 272: 1997-2004.
36 Michelson AD, Barnard MR. Flow cytometric detection of the redistribution of the glycoprotein Ib-IX complex on thrombin-stimulated platelets is dependent on the type of antibody conjugate used. Blood 1993; 81: 1408-9.
37 LaRosa CA, Rohrer MJ, Benoit SE, Barnard MR, Michelson AD. Neutrophil cathepsin G modulates the platelet surface expression of the glycoprotein (GP) Ib-IX complex by proteolysis of the von Willebrand factor binding site on GPIbα and by a cytoskeletal-mediated redistribution of the remainder of the complex. Blood 1994; 84: 158-68.
38 Molino M, Di Lallo M, Martelli N, de Gaetano G, Cerletti C. Effects of leukocyte-derived cathepsin G on platelet membrane glycoprotein Ib-IX and IIb-IIIa complexes: a comparison with thrombin. Blood 1993; 82: 2442-51.
39 Pidard D, Renesto P, Berndt MC, Rabhi S, Clemetson KJ, Chignard M. Neutrophil proteinase cathepsin G is proteolytically active on the human platelet glycoprotein Ib-IX receptor: characterization of the cleavage sites within the glycoprotein Ib alpha subunit. Biochem J 1994; 303: 489-98.
40 Michelson AD, Ellis PA, Barnard MR, Matic GB, Viles AF, Kestin AS. Downregulation of the platelet surface glycoprotein Ib-IX complex in whole blood stimulated by thrombin, ADP or an in vivo wound. Blood 1991; 77: 770-9.
41 Lu H, Menashi S, Garcia I. et al. Reversibility of thrombin-induced decrease in platelet glycoprotein Ib function. Br J Haematol 1993; 85: 116-23.
42 Kovacsovics TJ, Hartwig JH. Thrombin-induced GPIb-IX centralization on the platelet surface requires actin assembly and myosin II activation. Blood 1996; 87: 618-29.
43 Brower MS, Levin RI, Garry K. Human neutrophil elastase modulates platelet function by limited proteolysis of membrane glycoproteins. J Clin Invest 1985; 75: 657-66.
44 Wicki AN, Clemetson KJ. Structure and function of platelet membrane glycoproteins Ib and V. Effects of leukocyte elastase and other proteases on platelets response to von Willebrand factor and thrombin. Eur J Biochem 1985; 153: 1-11.
45 Ramachandran R, Klufas AS, Molino M, Ahuja M, Hoxie JA, Brass LF. Release of the thrombin receptor (PAR-1) N-terminus from the surface of human platelets activated by thrombin. Thromb Haemost 1997; 78: 1119-24.
46 Mazzucato M, Marco LD, Masotti A, Pradella P, Bahou WF, Ruggeri ZM. Characterization of the initial alpha-thrombin interaction with glycoprotein Ib alpha in relation to platelet activation. J Biol Chem 1998; 273: 1880-7.
47 Kahn ML, Zheng YW, Huang W. et al. A dual thrombin receptor system for platelet activation. Nature 1998; 394: 690-4.
48 Berndt MC, Phillips DR. Platelet membrane proteins: composition and receptor function. In: Gordon JL. ed. Platelets in Biology and Pathology. Amsterdam: Elsevier/North Holland Biomedical Press; 1981: 43-75.
49 Jamieson GA. Pathophysiology of platelet thrombin receptors. Thromb Haemost 1997; 78: 242-6.
50 Phillips DR, Agin PP. Platelet plasma membrane glycoproteins. Identification of a proteolytic substrate for thrombin. Biochem Biophys Res Commun 1977; 75: 940-7.
51 Brass LF, Vassallo Jr RR, Belmonte E, Ahuja M, Cichowski K, Hoxie JA. Structure and function of the human platelet thrombin receptor. Studies using monoclonal antibodies directed against a defined domain within the receptor N terminus. J Biol Chem 1992; 267: 13795-8.
52 Molino M, Bainton DF, Hoxie JA, Coughlin SR, Brass LF. Thrombin receptors on human platelets. Initial localization and subsequent redistribution during platelet activation. J Biol Chem 1997; 272: 6011-7.
53 Nelken NA, Soifer SJ, O’Keefe J, Vu TK, Charo IF, Coughlin SR. Thrombin receptor expression in normal and atherosclerotic human arteries. J Clin Invest 1992; 90: 4-21.