γA/γ’ fibrinogen inhibits thrombin-induced platelet aggregation

Rehana S. Lovely; Chantelle M. Rein; Tara C. White; Sari A. Jouihan; Lynn K. Boshkov; Antony C. Bakke; Owen J. McCarty; David H. Farrell

doi:10.1160/TH08-03-0145

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2008; 100(05): 837-846
DOI: 10.1160/TH08-03-0145

Platelets and Blood Cells

Schattauer GmbH

γA/γ’ fibrinogen inhibits thrombin-induced platelet aggregation

Rehana S. Lovely

¹Department of Biomedical Sciences, Missouri State University, Springfield, Missouri, USA

,

Chantelle M. Rein

²Department of Pathology

,

Tara C. White

³Division of Biomedical Engineering, Department of Science and Engineering, School of Medicine, Oregon Health & Science University, Portland, Oregon, USA

,

Sari A. Jouihan

²Department of Pathology

,

Lynn K. Boshkov

²Department of Pathology

,

Antony C. Bakke

²Department of Pathology

,

Owen J. McCarty

³Division of Biomedical Engineering, Department of Science and Engineering, School of Medicine, Oregon Health & Science University, Portland, Oregon, USA

,

David H. Farrell

²Department of Pathology

› Institutsangaben

Abstract

Summary

The minor γA/γ’ fibrinogen isoform contains a high affinity binding site for thrombin exosite II that is lacking in the major γA/γA fibrinogen isoform. We therefore investigated the biological consequences of the γ’ chain binding to thrombin. Thrombin-induced platelet aggregation was inhibited by γA/γ’ fibrinogen.Carboxyl terminal peptide fragment γ’410–427 from the γ’ chain was also inhibitory, with an IC₅₀ of ∼200 µM in whole plasma. Deletion of the peptide from either the amino or carboxyl end significantly decreased inhibition. In contrast to thrombin-induced platelet aggregation, aggregation induced by epinephrine, ADP, arachidonic acid, or SFLLRN peptide showed little inhibition by the γ’ peptide. The inhibition of thrombin-induced platelet aggregation was not due to direct inhibition of the thrombin active site, since cleavage of a small peptidyl substrate was 91% of normal even in the presence of 1 mM γ’410–427.The γ’410–427 peptide blocked platelet adhesion to immobilized thrombin under both static and flow conditions,blocked soluble thrombin binding to platelet GPIbα, and inhibited PAR1 cleavage by thrombin. These results suggest that the γ’ chain of fibrinogen inhibits thrombin-induced platelet aggregation by binding to thrombin exosite II. Thrombin that is bound to the γ’ chain is thereby prevented from activating platelets, while retaining its amidolytic activity.

Keywords

Antiplatelet agents - antithrombin - fibrinogen/fibrin - protease activated receptors (PAR) - thrombin

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Referenzen

References
1 Lord ST. Fibrinogen and fibrin: scaffold proteins in hemostasis. Curr Opin Hematol 2007; 14: 236-241.
2 Henschen-Edman AH. Fibrinogen non-inherited heterogeneity and its relationship to function in health and disease. Ann NY Acad Sci 2001; 936: 580-593.
3 Pan Y, Doolittle RF. cDNA sequence of a second fibrinogen a chain in lamprey: an archetypal version alignable with full-length β and γ chains. Proc Natl Acad Sci USA 1992; 89: 2066-2070.
4 Fu Y, Weissbach L, Plant PW. et al. Carboxy-terminal-extended variant of the human fibrinogen a subunit: a novel exon conferring marked homology to β and γ subunits. Biochemistry 1992; 31: 11968-11972.
5 Chung DW, Davie EW. γ and γ’ chains of human fibrinogen are produced by alternative mRNA processing. Biochemistry 1984; 23: 4232-4236.
6 Fornace AJ, Cummings DE, Comeau CM. et al. Structure of the human γ-fibrinogen gene, alternate mRNA splicing near the 3’ end of the gene produces γA and γB forms of γ-fibrinogen. J Biol Chem 1984; 259: 12826-12830.
7 Wolfenstein-Todel C, Mosesson MW. Human plasma fibrinogen heterogeneity: evidence for an extended carboxyl-terminal sequence in a normal γ chain variant (γ’). Proc Natl Acad Sci U S A 1980; 77: 5069-5073.
8 Legrele CD, Wolfenstein-Todel C, Hurbourg Y. et al. Evidence for two classes of rat plasma fibrinogen γ chains differing by their COOH-terminal amino acid sequences. BiochemBiophys Res Commun 1982; 105: 521-529.
9 Lovely RS, Falls LA, Al-Mondhiry HA. et al. Association of γA/γ fibrinogen levels and coronary artery disease. Thromb Haemost 2002; 88: 26-31.
10 Farrell DH, Thiagarajan P, Chung DW. et al. Role of fibrinogen α and γ chain sites in platelet aggregation. Proc Natl Acad Sci USA 1992; 89: 10729-10732.
11 Kirschbaum NE, Mosesson MW, Amrani DL. Characterization of the γ chain platelet binding site on fibrinogen fragment D. Blood 1992; 79: 2643-2648.
12 Lawrence SO, Wright TW, Francis CW. et al. Purification and characterization of homodimeric γB-γB fibrinogen from rat plasma. Blood 1993; 82: 2406-2413.
13 Farrell DH, Thiagarajan P. Binding of recombinant fibrinogen mutants to platelets. J Biol Chem 1994; 269: 226-231.
14 Siebenlist KR, Meh DA, Mosesson MW. Plasma factor XIII binds specifically to fibrinogen molecules containing γ’ chains. Biochemistry 1996; 35: 10448-10453.
15 Moaddel M, Farrell DH, Daugherty MA. et al. Interactions of human fibrinogens with factor XIII, roles of calcium and the γ’ peptide. Biochemistry 2000; 39: 6698-6705.
16 Meh DA, Siebenlist KR, Mosesson MW. Identification and characterization of the thrombin binding sites on fibrin. J Biol Chem 1996; 271: 23121-23125.
17 Meh DA, Siebenlist KR, Brennan SO. et al. The amino acid sequence in fibrin responsible for high affinity thrombin binding. Thromb Haemost 2001; 85: 470-474.
18 Lovely RS, Moaddel M, Farrell DH. Fibrinogen γ’ chain binds thrombin exosite II. J Thromb Haemost 2003; 01: 124-131.
19 Fredenburgh JC, Stafford AR, Leslie BA. et al. Bivalent binding to γA/γ’-fibrin engages both exosites of thrombin and protects it from inhibition by the anti thrombinheparin complex. J Biol Chem 2008; 283: 2470-2477.
20 Sabo TM, Farrell DH, Maurer MC. Conformational analysis of g’ peptide (410-427) interactions with thrombin anion binding exosite II. Biochemistry 2006; 45: 7434-7445.
21 Pineda AO, Chen ZW, Marino F. et al. Crystal structure of thrombin in complex with fibrinogen γ’ peptide. Biophys Chem 2007; 125: 556-559.
22 Falls LA, Farrell DH. Resistance of γA/γ’ fibrin clots to fibrinolysis. J Biol Chem 1997; 272: 14251-14256.
23 Collet JP, Nagaswami C, Farrell DH. et al. Influence of γ’ fibrinogen splice variant on fibrin physical properties and fibrinolysis rate. Arterioscler Thromb Vasc Biol 2004; 24: 382-286.
24 Siebenlist KR, Mosesson MW, Hernandez I. et al. Studies on the basis for the properties of fibrin produced from fibrinogen-containing γ’ chains. Blood 2005; 106: 2730-2736.
25 Weiler H. Mouse models of thrombosis: thrombomodulin. Thromb Haemost 2004; 92: 467-477.
26 Celikel R, McClintock RA, Roberts JR. et al. Modulation of α-thrombin function by distinct interactions with platelet glycoprotein Ibα. Science 2003; 301: 218-221.
27 Dumas JJ, Kumar R, Seehra J. et al. Crystal structure of the GPIbα-thrombin complex essential for platelet aggregation. Science 2003; 301: 222-226.
28 De Candia E, Hall SW, Rutella S. et al. Binding of thrombin to glycoprotein Ib accelerates the hydrolysis of PAR-1 on intact platelets. J Biol Chem 2001; 276: 4692-4698.
29 Li CQ, Vindigni A, Sadler JE. et al. Platelet glycoprotein Ibα binds to thrombin anion-binding exosite II inducing allosteric changes in the activity of thrombin. J Biol Chem 2001; 276: 6161-6168.
30 Pechik I, Madrazo J, Mosesson MW. et al. Crystal structure of the complex between thrombin and the central ‘E’ region of fibrin. Proc Natl Acad Sci USA 2004; 101: 2718-27123.
31 Lovely RS, Boshkov LK, Marzec UM. et al. Fibrinogen γ’ chain carboxy terminal peptide selectively inhibits the intrinsic coagulation pathway. Br J Haematol 2007; 139: 494-503.
32 de Bosch NB, Mosesson MW, Ruiz-Saez A. et al. Inhibition of thrombin generation in plasma by fibrin formation (antithrombin I). Thromb Haemost 2002; 88: 253-258.
33 McCarty OJ, Calaminus SD, Berndt MC. et al. von Willebrand factor mediates platelet spreading through glycoprotein Ib and αIIbβ3 in the presence of botrocetin and ristocetin, respectively. J Thromb Haemost 2006; 04: 1367-1378.
34 White TC, Berny MA, Robinson DK. et al. The leech product saratin is a potent inhibitor of platelet integrin α2β1 and von Willebrand factor binding to collagen. FEBS J 2007; 274: 1481-1491.
35 Hawiger J. Adhesive ends of fibrinogen and its antiadhesive peptides, the end of a saga?. Semin Hematol 1995; 32: 99-109.
36 Peerschke EIB, Francis CW, Marder VJ. Fibrinogen binding to human blood platelets: effect of γ chain carboxyterminal structure and length. Blood 1986; 67: 385-390.
37 Amrani DL, Newman PJ, Meh D. et al. The role of fibrinogen Aα chains in ADP-induced platelet aggregation in the presence of fibrinogen molecules containing γ’chains. Blood 1988; 72: 919-924.
38 Haidaris PJ, Peerschke EIB, Marder VJ. et al. The C-terminal sequences of the γ^57.5 chain of human fibrinogen constitute a plasmin sensitive epitope that is exposed in crosslinked fibrin. Blood 1989; 74: 2437-2444.
39 Berny MA, White TC, Tucker EI. et al. Thrombin mutant W215A/E217A acts as a platelet GPIb antagonist. Arterioscler Thromb Vasc Biol 2008; 28: 329-334.
40 Brass LF, Pizarro S, Ahuja M. et al. Changes in the structure and function of the human thrombin receptor during receptor activation, internalization, and recycling. J Biol Chem 1994; 269: 2943-2952.
41 Seegers WH, Nieft M, Loomis EC. Note on the adsorption of thrombin on fibrin. Science 1945; 101: 520-521.
42 Mosesson MW. Antithrombin I. Inhibition of thrombin generation in plasma by fibrin formation. Thromb Haemost 2003; 89: 9-12.
43 Francis CW, Nachman RL, Marder VJ. Plasma and platelet fibrinogen differ in γ chain content. Thromb Haemost 1984; 51: 84-88.
44 Mosesson MW, Homandberg GA, Amrani DL. Human platelet fibrinogen γ chain structure. Blood 1984; 63: 990-995.
45 Drouet L, Paolucci F, Pasqualini N. et al. Plasma γ’/γ fibrinogen ratio, a marker of arterial thrombotic activity: a new potential cardiovascular risk factor?. Blood Coagul Fibrinolysis 1999; 10 (Suppl. 01) S35-S39.
46 Mannila MN, Lovely RS, Kazmierczak SC. et al. Elevated plasma fibrinogen γ’ concentration is associated with myocardial infarction: effects of variation in fibrinogen genes and environmental factors. J Thromb Haemost 2007; 05: 766-773.
47 Cheung EY, de Willige SU, Vos HL. et al. Fibrinogen γ’ in ischemic stroke: a case-control study. Stroke 2008; 39: 1033-1035.
48 Mosesson MW, Hernandez I, Raife TJ. et al. Plasma fibrinogen γ’ chain content in the thrombotic microangiopathy syndrome. J Thromb Haemost 2007; 05: 62-69.
49 Uitte de Willige S, de Visser MC, Houwing-Duistermaat JJ. et al. Genetic variation in the fibrinogen g gene increases the risk for deep venous thrombosis by reducing plasma fibrinogen γ’ levels. Blood 2005; 106: 4176-4183.
50 Mann KG, Brummel-Ziedins K, Orfeo T. et al. Models of blood coagulation. Blood Cells Mol Dis 2006; 36: 108-117.