Download PDF
Thromb Haemost 2023; 123(04): 380-392
DOI: 10.1055/a-1993-4193
Coagulation and Fibrinolysis

Transglutaminase Activities of Blood Coagulant Factor XIII Are Dependent on the Activation Pathways and on the Substrates

Rameesa D. Syed Mohammed
1   Department of Chemistry, University of Louisville, Louisville, Kentucky, United States
,
Francis D. O. Ablan
1   Department of Chemistry, University of Louisville, Louisville, Kentucky, United States
,
Nicholas M. McCann
1   Department of Chemistry, University of Louisville, Louisville, Kentucky, United States
,
Mohammed M. Hindi
1   Department of Chemistry, University of Louisville, Louisville, Kentucky, United States
,
Muriel C. Maurer
1   Department of Chemistry, University of Louisville, Louisville, Kentucky, United States
› Author Affiliations Funding This research was supported by a grant from the National Institutes of Health (R15 HL120068).
› Further Information
    Permissions and Reprints


Abstract

Factor XIII (FXIII) catalyzes formation of γ-glutamyl-ε-lysyl crosslinks between reactive glutamines (Q) and lysines (K). In plasma, FXIII is activated proteolytically (FXIII-A*) by the concerted action of thrombin and Ca2+. Cellular FXIII is activated nonproteolytically (FXIII-A°) by elevation of physiological Ca2+ concentrations. FXIII-A targets plasmatic and cellular substrates, but questions remain on correlating FXIII activation, resultant conformational changes, and crosslinking function to different physiological substrates. To address these issues, the characteristics of FXIII-A* versus FXIII-A° that contribute to transglutaminase activity and substrate specificities were investigated. Crosslinking of lysine mimics into a series of Q-containing substrates were measured using in-gel fluorescence, mass spectrometry, and UV-Vis spectroscopy. Covalent incorporation of fluorescent monodansylcadaverine revealed that FXIII-A* exhibits greater activity than FXIII-A° toward Q residues within Fbg αC (233–425 WT, Q328P Seoul II, and Q328PQ366N) and actin. FXIII-A* and FXIII-A° displayed similar activities toward α2-antiplasmin (α2AP), fibronectin, and Fbg αC (233–388, missing FXIII-binding site αC 389–402). Furthermore, the N-terminal α2AP peptide (1–15) exhibited similar kinetic properties for FXIII-A* and FXIII-A°. MALDI-TOF mass spectrometry assays with glycine ethyl ester and Fbg αC (233–425 WT, αC E396A, and truncated αC (233–388) further documented that FXIII-A* exerts greater benefit from the αC 389–402 binding site than FXIII-A°. Conformational properties of FXIII-A* versus A° are proposed to help promote transglutaminase function toward different substrates. A combination of protein substrate disorder and secondary FXIII-binding site exposure are utilized to control activity and specificity. From these studies, greater understandings of how FXIII-A targets different substrates are achieved.

Keywords

antiplasmin - factor XIII - fibrinogen - fibronectin - actin

Authors Contribution

R.D.S.M, F.D.O.A., and M.C.M. designed the research. R.D.S.M., F.D.O.A., N.M.M., and M.M.H. expressed the proteins and carried out the experiments. R.D.S.M. performed the MDC assays. F.D.O.A., N.M.M., and M.M.H. performed the GEE Mass Spectrometry assays. R.D.S.M. and N.M.M. contributed to the PONDR analysis. M.C.M., R.D.S.M., and F.D.O.A. interpreted the data and wrote the manuscript. All authors reviewed the manuscript and approved its final version.


Supplementary Material



Publication History

Received: 02 August 2022

Accepted: 04 December 2022

Accepted Manuscript online:
06 December 2022

Article published online:
25 January 2023

© 2023. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Richardson VR, Cordell P, Standeven KF, Carter AM. Substrates of factor XIII-A: roles in thrombosis and wound healing. Clin Sci (Lond) 2013; 124 (03) 123-137
    CrossrefPubMedGoogle Scholar
  • 2 Schroeder V, Kohler HP. Factor XIII: structure and function. Semin Thromb Hemost 2016; 42 (04) 422-428
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Muszbek L, Bereczky Z, Bagoly Z, Komáromi I, Katona É. Factor XIII: a coagulation factor with multiple plasmatic and cellular functions. Physiol Rev 2011; 91 (03) 931-972
    CrossrefPubMedGoogle Scholar
  • 4 Byrnes JR, Wolberg AS. Newly-recognized roles of factor XIII in thrombosis. Semin Thromb Hemost 2016; 42 (04) 445-454
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Mitchell JL, Mutch NJ. Let's cross-link: diverse functions of the promiscuous cellular transglutaminase factor XIII-A. J Thromb Haemost 2019; 17 (01) 19-30
    CrossrefPubMedGoogle Scholar
  • 6 Alshehri FSM, Whyte CS, Mutch NJ. Factor XIII-A: an indispensable “factor” in haemostasis and wound healing. Int J Mol Sci 2021; 22 (06) 3055
    CrossrefPubMedGoogle Scholar
  • 7 Lorand L. Factor XIII: structure, activation, and interactions with fibrinogen and fibrin. Ann N Y Acad Sci 2001; 936: 291-311
    CrossrefPubMedGoogle Scholar
  • 8 Polgár J, Hidasi V, Muszbek L. Non-proteolytic activation of cellular protransglutaminase (placenta macrophage factor XIII). Biochem J 1990; 267 (02) 557-560
    CrossrefPubMedGoogle Scholar
  • 9 Hornyak TJ, Bishop PD, Shafer JA. Alpha-thrombin-catalyzed activation of human platelet factor XIII: relationship between proteolysis and factor XIIIa activity. Biochemistry 1989; 28 (18) 7326-7332
    CrossrefPubMedGoogle Scholar
  • 10 Standeven KF, Carter AM, Grant PJ. et al. Functional analysis of fibrin gamma-chain cross-linking by activated factor XIII: determination of a cross-linking pattern that maximizes clot stiffness. Blood 2007; 110 (03) 902-907
    CrossrefPubMedGoogle Scholar
  • 11 Helms CC, Ariëns RA, Uitte de Willige S, Standeven KF, Guthold M. α-α Cross-links increase fibrin fiber elasticity and stiffness. Biophys J 2012; 102 (01) 168-175
    CrossrefPubMedGoogle Scholar
  • 12 Byrnes JR, Duval C, Wang Y. et al. Factor XIIIa-dependent retention of red blood cells in clots is mediated by fibrin α-chain crosslinking. Blood 2015; 126 (16) 1940-1948
    CrossrefPubMedGoogle Scholar
  • 13 Sakata Y, Aoki N. Cross-linking of alpha 2-plasmin inhibitor to fibrin by fibrin-stabilizing factor. J Clin Invest 1980; 65 (02) 290-297
    CrossrefPubMedGoogle Scholar
  • 14 Lee KN, Lee CS, Tae WC, Jackson KW, Christiansen VJ, McKee PA. Cross-linking of wild-type and mutant alpha 2-antiplasmins to fibrin by activated factor XIII and by a tissue transglutaminase. J Biol Chem 2000; 275 (48) 37382-37389
    CrossrefPubMedGoogle Scholar
  • 15 Matsuka YV, Migliorini MM, Ingham KC. Cross-linking of fibronectin to C-terminal fragments of the fibrinogen alpha-chain by factor XIIIa. J Protein Chem 1997; 16 (08) 739-745
    CrossrefPubMedGoogle Scholar
  • 16 Alshehri FSM, Whyte CS, Tuncay A, Williams ML, Wilson HM, Mutch NJ. Monocytes expose factor XIII-A and stabilize thrombi against fibrinolytic degradation. Int J Mol Sci 2021; 22 (12) 6591
    CrossrefPubMedGoogle Scholar
  • 17 Mitchell JL, Lionikiene AS, Fraser SR, Whyte CS, Booth NA, Mutch NJ. Functional factor XIII-A is exposed on the stimulated platelet surface. Blood 2014; 124 (26) 3982-3990
    CrossrefPubMedGoogle Scholar
  • 18 Somodi L, Beke Debreceni I, Kis G. et al. Activation mechanism dependent surface exposure of cellular factor XIII on activated platelets and platelet microparticles. J Thromb Haemost 2022; 20 (05) 1223-1235
    CrossrefPubMedGoogle Scholar
  • 19 Mousa A, Cui C, Song A. et al. Transglutaminases factor XIII-A and TG2 regulate resorption, adipogenesis and plasma fibronectin homeostasis in bone and bone marrow. Cell Death Differ 2017; 24 (05) 844-854
    CrossrefPubMedGoogle Scholar
  • 20 Tsurupa G, Tsonev L, Medved L. Structural organization of the fibrin(ogen) alpha C-domain. Biochemistry 2002; 41 (20) 6449-6459
    CrossrefPubMedGoogle Scholar
  • 21 Duval C, Allan P, Connell SD, Ridger VC, Philippou H, Ariëns RA. Roles of fibrin α- and γ-chain specific cross-linking by FXIIIa in fibrin structure and function. Thromb Haemost 2014; 111 (05) 842-850
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 McPherson HR, Duval C, Baker SR. et al. Fibrinogen αC-subregions critically contribute blood clot fibre growth, mechanical stability, and resistance to fibrinolysis. eLife 2021; 10: 10
    CrossrefPubMedGoogle Scholar
  • 23 Hur WS, Paul DS, Bouck EG. et al. Hypofibrinogenemia with preserved hemostasis and protection from thrombosis in mice with an Fga truncation mutation. Blood 2022; 139 (09) 1374-1388
    CrossrefPubMedGoogle Scholar
  • 24 Mouapi KN, Bell JD, Smith KA, Ariëns RA, Philippou H, Maurer MC. Ranking reactive glutamines in the fibrinogen αC region that are targeted by blood coagulant factor XIII. Blood 2016; 127 (18) 2241-2248
    CrossrefPubMedGoogle Scholar
  • 25 Schmitt LR, Henderson R, Barrett A. et al. Mass spectrometry-based molecular mapping of native FXIIIa cross-links in insoluble fibrin clots. J Biol Chem 2019; 294 (22) 8773-8778
    CrossrefPubMedGoogle Scholar
  • 26 Smith KA, Adamson PJ, Pease RJ. et al. Interactions between factor XIII and the alphaC region of fibrinogen. Blood 2011; 117 (12) 3460-3468
    CrossrefPubMedGoogle Scholar
  • 27 Smith KA, Pease RJ, Avery CA. et al. The activation peptide cleft exposed by thrombin cleavage of FXIII-A(2) contains a recognition site for the fibrinogen α chain. Blood 2013; 121 (11) 2117-2126
    CrossrefPubMedGoogle Scholar
  • 28 Mouapi KN, Wagner LJ, Stephens CA. et al. Evaluating the effects of fibrinogen αC mutations on the ability of factor XIII to crosslink the reactive αC glutamines (Q237, Q328, Q366). Thromb Haemost 2019; 119 (07) 1048-1057
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Anokhin BA, Stribinskis V, Dean WL, Maurer MC. Activation of factor XIII is accompanied by a change in oligomerization state. FEBS J 2017; 284 (22) 3849-3861
    CrossrefPubMedGoogle Scholar
  • 30 Anokhin BA, Dean WL, Smith KA. et al. Proteolytic and nonproteolytic activation mechanisms result in conformationally and functionally different forms of coagulation factor XIII A. FEBS J 2020; 287 (03) 452-464
    CrossrefPubMedGoogle Scholar
  • 31 Ayala YM, Cantwell AM, Rose T, Bush LA, Arosio D, Di Cera E. Molecular mapping of thrombin-receptor interactions. Proteins 2001; 45 (02) 107-116
    CrossrefPubMedGoogle Scholar
  • 32 Murrey HE, Judkins JC, Am Ende CW. et al. Systematic evaluation of bioorthogonal reactions in live cells with clickable HaloTag ligands: implications for intracellular imaging. J Am Chem Soc 2015; 137 (35) 11461-11475
    CrossrefPubMedGoogle Scholar
  • 33 Doiphode PG, Malovichko MV, Mouapi KN, Maurer MC. Evaluating factor XIII specificity for glutamine-containing substrates using a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry assay. Anal Biochem 2014; 457: 74-84
    CrossrefPubMedGoogle Scholar
  • 34 Park R, Doh HJ, An SS, Choi JR, Chung KH, Song KS. A novel fibrinogen variant (fibrinogen Seoul II; AalphaGln328Pro) characterized by impaired fibrin alpha-chain cross-linking. Blood 2006; 108 (06) 1919-1924
    CrossrefPubMedGoogle Scholar
  • 35 To WS, Midwood KS. Plasma and cellular fibronectin: distinct and independent functions during tissue repair. Fibrogenesis Tissue Repair 2011; 4: 21
    CrossrefPubMedGoogle Scholar
  • 36 Bender M, Palankar R. Platelet shape changes during thrombus formation: role of actin-based protrusions. Hamostaseologie 2021; 41 (01) 14-21
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Cohen I, Blankenberg TA, Borden D, Kahn DR, Veis A. Factor XIIIa-catalyzed cross-linking of platelet and muscle actin. Regulation by nucleotides. Biochim Biophys Acta 1980; 628 (03) 365-375
    CrossrefPubMedGoogle Scholar
  • 38 Hoffmann BR, Annis DS, Mosher DF. Reactivity of the N-terminal region of fibronectin protein to transglutaminase 2 and factor XIIIA. J Biol Chem 2011; 286 (37) 32220-32230
    CrossrefPubMedGoogle Scholar
  • 39 Lefebvre P, Velasco PT, Dear A. et al. Severe hypodysfibrinogenemia in compound heterozygotes of the fibrinogen AalphaIVS4 + 1G>T mutation and an AalphaGln328 truncation (fibrinogen Keokuk). Blood 2004; 103 (07) 2571-2576
    CrossrefPubMedGoogle Scholar
  • 40 Thomas A, Biswas A, Dodt J. et al. Coagulation factor XIIIA subunit missense mutations affect structure and function at the various steps of factor XIII action. Hum Mutat 2016; 37 (10) 1030-1041
    CrossrefPubMedGoogle Scholar
  • 41 Schroeder V, Vuissoz JM, Caflisch A, Kohler HP. Factor XIII activation peptide is released into plasma upon cleavage by thrombin and shows a different structure compared to its bound form. Thromb Haemost 2007; 97 (06) 890-898
    Article in Thieme ConnectPubMedGoogle Scholar
  • 42 Stieler M, Weber J, Hils M. et al. Structure of active coagulation factor XIII triggered by calcium binding: basis for the design of next-generation anticoagulants. Angew Chem Int Ed Engl 2013; 52 (45) 11930-11934
    CrossrefPubMedGoogle Scholar
  • 43 Woofter RT, Maurer MC. Role of calcium in the conformational dynamics of factor XIII activation examined by hydrogen-deuterium exchange coupled with MALDI-TOF MS. Arch Biochem Biophys 2011; 512 (01) 87-95
    CrossrefPubMedGoogle Scholar
  • 44 Hornyak TJ, Shafer JA. Interactions of factor XIII with fibrin as substrate and cofactor. Biochemistry 1992; 31 (02) 423-429
    CrossrefPubMedGoogle Scholar
  • 45 Kattula S, Bagoly Z, Tóth NK, Muszbek L, Wolberg AS. The factor XIII-A Val34Leu polymorphism decreases whole blood clot mass at high fibrinogen concentrations. J Thromb Haemost 2020; 18 (04) 885-894
    CrossrefPubMedGoogle Scholar
  • 46 Trumbo TA, Maurer MC. Examining thrombin hydrolysis of the factor XIII activation peptide segment leads to a proposal for explaining the cardioprotective effects observed with the factor XIII V34L mutation. J Biol Chem 2000; 275 (27) 20627-20631
    CrossrefPubMedGoogle Scholar
  • 47 Li B, Billur R, Maurer MC. et al. Proline 36 of the factor XIII activation peptide plays a crucial role in substrate recognition and zymogen activation. Thromb Haemost 2018; 118 (12) 2037-2045
    Article in Thieme ConnectPubMedGoogle Scholar
  • 48 Tsurupa G, Yakovlev S, McKee P, Medved L. Noncovalent interaction of alpha(2)-antiplasmin with fibrin(ogen): localization of alpha(2)-antiplasmin-binding sites. Biochemistry 2010; 49 (35) 7643-7651
    CrossrefPubMedGoogle Scholar
  • 49 Makogonenko E, Tsurupa G, Ingham K, Medved L. Interaction of fibrin(ogen) with fibronectin: further characterization and localization of the fibronectin-binding site. Biochemistry 2002; 41 (25) 7907-7913
    CrossrefPubMedGoogle Scholar
  • 50 Serrano K, Devine DV. Intracellular factor XIII crosslinks platelet cytoskeletal elements upon platelet activation. Thromb Haemost 2002; 88 (02) 315-320
    Article in Thieme ConnectPubMedGoogle Scholar