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2015 | 62 | 3 | 517-522
Article title

Non-enzymatic activation of prothrombin induced by interaction with fibrin β26-42 region

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We have discovered that addition of monomeric desAB fibrin to prothrombin leads to appearance of the thrombin-like activity of prothrombin towards S2238 chromogenic substrate. DesA and desABβ(15-42)2 fibrin forms did not cause any activation of prothrombin. From this observation we could suggested that amino acid residues of the 15-42 fragment of BβN-domain presented in desAB fibrin, cleaved in desABβ(15-42)2 fibrin and protected in desA fibrin, play a crucial role in the non-enzymatic activation of prothrombin. To identify the Bβ amino acid residues involved in the fibrin-prothrombin binding we used monoclonal antibodies 1-5G and 2d2a with epitopes in Bβ26-42 and Bβ12-26 fibrin fragments respectively. The thrombin-like activity in the mixture of prothrombin and desAB fibrin was monitored in the presence of each of these monoclonal antibodies. It was found that anti-Bβ12-26 antibody does not exhibit any inhibitory effect on the thombin-like activity of the mixture. In contrast, adding of Bβ26-42 antibody into the mixture of desAB fibrin with prothrombin diminished the thrombin-like activity by 70%. Recombinant dimeric peptides Bβ(15-44)2 and Bβ(15-66)2 that mimic amino acid residues in fibrin were also tested for their ability to activate prothrombin. It was found that both peptides were able to induce non-enzymatic activation of prothrombin. The activation was more evident in the case of Bβ(15-44)2 peptide. From the data obtained we can conclude that desAB fibrin binds to prothrombin through the Bβ26-42 amino acid residues and the formation of such a complex caused a non-enzymatic activation of prothrombin.
Physical description
  • Belitser VA, Pozdnjakova TM, Platonova TN, Vovk EV (1981) Fibrin-fragment D complex formation. Thromb Res 21: 565-572.
  • Bennet JS (2001) Platelet-fibrinogen interactions. Ann N Y Acad Sci 936: 340-354.
  • Binnie CG, Lord ST (1991) A synthetic analog of fibrinogen alpha 27-50 is an inhibitor of thrombin. Thromb Haemost 65: 165-168.
  • Binnie CG, Lord ST (1993) The fibrinogen sequences that interact with thrombin. Blood 81: 3186-3192.
  • Bradford HN, Micucci JA, Krishnaswamy S (2010) Regulated cleavage of prothrombin by prothrombinase. J Biol Chem 285: 328-338.
  • Chernyshenko VO, Gornytska OV., Platonova TM, Sokolovska LI (2010) A new fibrinogenase from Echis multisquamatis venom is a perspective agent for limited proteolysis and defibrinogenation. ABB 1: 91-96.
  • Chernyshenko VO, Platonova TM, Makogonenko YM, Rebriev AV, Mikhalovska LI, Chernyshenko TM, Komisarenko SV (2014) Fibrin(ogen)olytic and platelet modulating activity of a novel protease from the Echis multisquamatis snake venom. Biochimie 105: 76-83.
  • Di Cera E (2003) Thrombin interactions. Chest 124: 11S-17S.
  • DiBella EE, Scheraga HA (1996) The role of the insertion loop around tryptophan 148 in the activity of thrombin. Biochemistry 35: 4427-4433.
  • Friedrich R, Panizzi P, Fuentes-Prior P (2003) Staphylocoagulase is a prototype for the mechanism of cofactorinduced zymogen activation. Nature 425: 535-540.
  • Friedrich R, Panizzi P, Kawabata S (2006) Structural basis for reduced staphylocoagulase-mediated bovine prothrombin activation. J Biol Chem 281: 1188-1195.
  • Gershkovich AA, Kibirev VK (1988) Chromogenic and fluorogenic peptide substrates of proteolytic enzymes. Bioorg Khim 14: 1461-1488.
  • Goodwin CA, Kakkar VV, Scully MF (1992) Generation of forms of fragment E with differing thrombin-binding properties during digestion of fibrinogen by plasmin. Biochem J 281: 613-618.
  • Gorlatov S, Medved L (2002) Interaction of fibrin(ogen) with the endothelial cell receptor VE-cadherin: mapping of the receptor-binding site in the NH2-terminal portions of the fibrin beta chains. Biochemistry 41: 4107-4116.
  • Gun'ko VM, Klyueva AV, Levchuk YN, Leboda R (2003) Photon correlation spectroscopy investigations of proteins. Adv Colloid Interface Sci 105: 201-328.
  • Hogg PJ, Jackson CM, Labanowski JK, Bock PE (1996) Binding of fibrin monomer and heparin to thrombin in a ternary complex alters the environment of the thrombin catalytic site, reduces affinity for hirudin, and inhibits cleavage of fibrinogen. Biol Chem 271: 26088-26095.
  • Kaczmarek E, Kaminski M, McDonagh J (1987) Fibrinogen-sepharose interaction with prothrombin, prethrombin 1, prethrombin 2 and thrombin (1987) Biochim Biophys Acta 914: 275-282.
  • Kaczmarek E, McDonagh J (1988) Thrombin binding to the A alpha-, B beta-, and gamma-chains of fibrinogen and to their remnants contained in fragment E. J Biol Chem 263: 13896-13900.
  • Khan AR, James MN (1998) Molecular mechanisms for the conversion of zymogens to active proteolytic enzymes. Protein Science 7: 815-836.
  • Lane DA, Philippou H, Huntington JA (2005) Directing thrombin. Blood 106: 2605-2612.
  • Lorber B, Fischer F, Bailly M (2012) Protein analysis by dynamic light scattering: methods and techniques for students. Biochem Mol Biol Educ 40: 372-382.
  • Lugovskoy EV, Gritsenko PG, Kapustianenko LG, Kolesnikova IN, Chernishov VI, Komisarenko SV (2007) Functional role of Bβ-chain N-terminal fragment in the fibrin polymerization process. FEBS J 274: 4540-4549.
  • Mann KG (1976) Prothrombin. Methods Enzymol 45: 123-156.
  • Medved LV, Platonova TN, Litvinovich SV, Lukinova NI (1988) The cleavage of beta-chain in bovine fibrinogen DH fragment (95 kDa) leads to a significant increase in its anticlotting activity. FEBS Lett 232: 56-60.
  • Meh DA, Siebenlist KR, Mosesson MW (1996) Identification and characterization of the thrombin binding sites on fibrin. J Biol Chem 271: 23121-23125.
  • Mosesson MW (1993) Thrombin interactions with fibrinogen and fibrin. Semin Thromb Hemost 19: 361-367.
  • Mosesson MW (2003) Fibrinogen gamma chain functions. J Thromb Haemost 1: 231-238.
  • Mosesson MW, Hernandez I, Siebenlist KR (2004) Evidence that catalytically-inactivated thrombin forms non-covalently linked dimers that bridge between fibrin/fibrinogen fibers and enhance fibrin polymerization. Biophys Chem 110: 93-100.
  • Ni F, Ning Q, Jackson CM, Fenton JW (1993) Thrombin exosite for fibrinogen recognition is partially accessible in prothrombin. J Biol Chem 268: 16899-16902.
  • Pierce BG, Wiehe K, Hwang H, Kim BH, Vreven T, Weng Z (2014) ZDOCK Server: Interactive Docking Prediction of Protein-Protein Complexes and Symmetric Multimers. Bioinformatics 30: 1771-1773.
  • Platonova TM, Slominsky AYu, Chernyshenko TM, Makogonenko EM (2002) The blood coagulation system key proenzymes activation by fibrin E-fragment. Ukr Biokhim Zh 74: 25-29.
  • Pospisil CH, Stafford AR, Fredenburgh JC, Weitz JI (2003) Evidence that both exosites on thrombin participate in its high affinity interaction with fibrin. J Biol Chem 278: 21584-21591.
  • Roy A, Kucukural A, Zhang Y (2010) I-TASSER: a unified platform for automated protein structure and function prediction. Nature Protocols 5: 725-738.
  • Roy A, Yang J, Zhang Y (2012) COFACTOR: An accurate comparative algorithm for structure-based protein function annotation. Nucleic Acids Research 40: W471-W477.
  • Savchuk OM, Krasnobryzha IM, Horchev VF, Chernyshenko TM, Havryliuk SP, Platonova TM (2006) Process of complexation of prothrombin and fibrin E-fragment. Ukr Biokhim Zh 78: 99-105.
  • Scheraga HA (2004) The thrombin-fibrinogen interaction. Biophys Chem 112: 117-130.
  • Solov'ev DA, Ugarova TP (1993) Isolation and characteristics of alpha-specific thrombin-like enzymes from venoms of the common pit viper (Agkistrodon halys halys) and the eastern pit viper (the central Asian subspecies Agkistrodon halys blomhoffii). Biokhimiia 58: 1221-1233.
  • van de Locht A, Stubbs MT, Bauer M, Bode W (1996) Crystallographic evidence that the F2 kringle catalytic domain linker of prothrombin does not cover the fibrinogen recognition exosite. J Biol Chem 271: 3413-3416.
  • Varetskaia TV (1965) Preparation of a fibrin monomer and studies on some of its properties. Ukr Biokhim Zh 37: 194-206.
  • Wolberg AS (2007) Thrombin generation and fibrin clot structure. Blood Rev 21: 131-142.
  • Yakovlev S, Gorlatov S, Ingham K, Medved L (2003) Interaction of Fibrin(ogen) with Heparin: Further Characterization and Localization of the Heparin-Binding Site Biochemistry 42: 7709-7716.
  • Yakovlev S, Medved L (2009) Interaction of fibrin(ogen) with endothelial cell receptor VE-Cadherin: localization of the fibrin-binding site within the third extracellular VE-cadherin domain. Biochemistry 48: 5171-5179.
  • Zhang Y (2008) I-TASSER server for protein 3D structure prediction. BMC Bioinformatics 9: 40.
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