Protransglutaminase (Factor XIII) Mediated Crosslinking of Fibrinogen and Fibrin*

Summary

Plasma factor XIII (plasma protransglutaminase) circulates as an A₂B₂ tetramer bound to the γ’ variant chains of fibrinogen “2”. During clotting the A subunits of fXIII are cleaved by thrombin to form fXIIIa (transglutaminase) and in the presence of calcium ions, activated A₂* subunits dissociate from the B subunits. When purified plasma fXIII or recombinant cellular factor XIII (A₂) was incubated with fibrinogen in the presence of calcium ions (≥50 μM) a non-synerizing gel formed concomitant with formation of γ dimers, followed by A γ polymers, and eventually γ trimers and γ tetramers. As is the case of fXIIIa, the fXIII-mediated crosslinking rate was enhanced in the presence of thiols. After an initial lag period, fXIII catalyzed fibrinogen cross-linking at ~75% of the rate of fXIIIa under typical crosslinking conditions (100 Loewy u/ml, 5 mM CaCl₂ & 500 μM DTT). Fibrin was crosslinked about 8 times more rapidly by fXIII than was fibrinogen, and after an initial lag period fXIII crosslinked fibrin at nearly the same rate as fXIIIa. Substituting plasma for purified fXIII as the source for fXIII resulted in robust fibrinogen crosslinking activity. In contrast to the high level of fXIII-mediated crosslinking activity observed with fibrinogen or fibrin as substrates, when transglutamination was measured using cadaverine incorporation into casein, fXIII was 30-fold less active than fXIIIa. Thus, factor XIII displays constitutive enzymatic activity with respect to fibrinogen and fibrin. The results further indicate that uncleaved fXIII in plasma provides a potent source of readily available crosslinking activity in clotting blood. Fibrinogen 2, whose γ’chains bind fXIII B subunits, was crosslinked 3.5 times more slowly by fXIII than was fibrinogen 1 (lacking γ’ chains), suggesting that complex formation between fibrinogen 2 and plasma fXIII plays a significant role in down-regulating potential plasma fXIII-mediated crosslinking activity. Since fibrin is a considerably better substrate for fXIII than is fibrinogen, the rate at which crosslinking takes place in a fibrinogen-containing plasma environment is much lower than it would be if fibrin were present.

Keywords

Factor XIII - fibrinogen - protransglutaminase - transglutaminase - familial mediterranean fever - α₂-antiplasmin

• References

• 1 Mosesson MW, Finlayson JS. The Search for the Structure of Fibrinogen. In: Progress in Hemostasis and Thrombosis. Spaet TH. ed. Orlando: Grune and Stratton; 1976. 3: 61.
• 2 Henschen A, Lottspeich F, Kehl M, Southan C. Covalent Structure of Fibrinogen. Ann NY Acad Sci 1983; 408: 28-43.
• 3 Doolittle RF. The Structure and Evolution of Vertebrate Fibrinogen. Ann NY Acad Sci 1983; 408: 13-27.
• 4 Finlayson JS, Mosesson MW. Heterogeneity of Human Fibrinogen. Biochemistry 1963; 2: 42-6.
• 5 Mosesson MW, Finlayson JS, Umfleet RA. Human Fibrinogen Heterogeneities III Identification of γ Chain Variants. J Biol Chem 1972; 247: 5223-327.
• 6 Mosesson MW, Finlayson JS, Umfleet RA, Galanakis D. Heterogeneities of Human Fibrinogen I Structural and Related Studies of Plasma Fibrinogens which are High Solubility Catabolic Intermediates. J Biol Chem 1972; 247: 5210-9.
• 7 Wolfenstein-Todel C, Mosesson MW. Human Plasma Fibrinogen Heterogeneity: Evidence for an Extended Carboxyl-Terminal Sequence in a Normal Gamma Chain Variant (γ’). Proc Natl Acad Sci USA 1980; 77: 5069-73.
• 8 Wolfenstein-Todel C, Mosesson MW. Carboxy-Terminal Amino Acid Sequence of a Human Fibrinogen Gamma-Chain Variant (γ’). Biochemistry 1981; 20: 6146-9.
• 9 Siebenlist KR, Meh DA, Mosesson MW. Plasma Factor XIII Binds Specifically to Fibrinogen Molecules Containing γ’ Chains. Biochemistry 1996; 35: 10448-53.
• 10 Meh DA, Siebenlist KR, Mosesson MW. Identification and Characterization of the Thrombin Binding Sites on Fibrin. J Biol Chem 1996; 271: 23121-5.
• 11 Kudryk B, Reuterby J, Blombäck B. Adsorption of Plasmic Fragment D to Thrombin Modified Fibrinogen-Sepharose. Thromb Res 1973; 2: 297-304.
• 12 Blombäck B, Hessel B, Hogg D, Therkildsen L. A Two-Step Fibrinogen-Fibrin Transition in Blood Coagulation. Nature 1978; 275: 501-5.
• 13 Olexa SA, Budzynski AZ. Evidence for Four Different Polymerization Sites Involved in Human Fibrin Formation. Proc Natl Acad Sci USA 1980; 77: 1374-8.
• 14 Olexa SA, Budzynski AZ, Doolittle RF, Cottrell BA, Greene TC. Structure of Fragment E Species from Human Crosslinked Fibrin. Biochemistry 1981; 21: 6139-45.
• 15 Budzynski AZ, Olexa SA, Pandya BV. Fibrin Polymerization Sites in Fibrinogen and Fibrin Fragments. Ann NY Acad Sci 1983; 408: 301-14.
• 16 Laudano AP, Cottrell BA, Doolittle RF. Synthetic Peptides Modeled on Fibrin Polymerization Sites. Ann NY Acad Sci 1983; 408: 315-29.
• 17 Shainoff JR, Dardik BN. Fibrinopeptide B and Aggregation of Fibrinogen. Science 1979; 204: 200-2.
• 18 Shainoff JR, Dardik BN. Fibrinopeptide B in Fibrin Assembly and Metabolism: Physiologic Significance in Delayed Release of the Peptide. Ann NY Acad Sci 1983; 408: 254-67.
• 19 Mosesson MW, Siebenlist KR, Amrani DL, DiOrio JP. Identification of Covalently Linked Trimeric and Tetrameric D Domains in Crosslinked Fibrin. Proc Natl Acad Sci USA 1989; 86: 1113-7.
• 20 Ferry JD. The Mechanism of Polymerization of Fibrinogen. Proc Natl Acad Sci USA 1952; 38: 566-9.
• 21 Stryer L, Cohen C, Langridge R. Axial Period of Fibrinogen and Fibrin. Nature 1963; 197: 793-4.
• 22 Krakow W, Endres GF, Siegel BM, Scheraga HA. An Electron Microscopic Investigation of the Polymerization of Bovine Fibrin Monomer. J Mol Biol 1972; 71: 95-103.
• 23 Hantgan RR, Hermans J. Assembly of Fibrin: A Light Scattering Study. J Biol Chem 1979; 254: 11272-81.
• 24 Fowler WE, Hantgan RR, Hermans J, Erickson HP. Structure of the Fibrin Protofibril. Proc Nat Acad Sci USA 1981; 78: 4872-6.
• 25 Williams RC. Morphology of Bovine Fibrinogen Monomers and Fibrin Oligomers. J Mol Biol 1981; 150: 399-408.
• 26 Williams RC. Morphology of Fibrinogen Monomers and of Fibrin Protofibrils. Ann NY Acad Sci 1983; 408: 180-93.
• 27 Carr Jr ME, Shen LL, Hermans J. Mass-Length Ratio of Fibrin Fibres From Gel Permeation and Light Scattering. Biopolymers 1977; 16: 1-15.
• 28 Hantgan R, Fowler W, Erickson H, Hermans J. Fibrin Assembly: A Comparison of Electron Microscopic and Light Scattering Results. Thromb Haemost 1980; 44: 119-24.
• 29 Pisano JJ, Finlayson JS, Peyton MP. Crosslink in Fibrin Polymerized by Factor XIII: ε-(γ-glutamyl)lysine. Science 1968; 160: 892-3.
• 30 Matacčić S, Loewy AG. The Identification fo Isopeptide Crosslinks in Insoluble Fibrin. Biochem Biophys Res Comm 1968; 30: 356-62.
• 31 Chen R, Doolittle RF. Identification of the Polypeptide Chains Involved in the Cross-linking of Fibrin. Proc Nat Acad Sci USA 1969; 63: 420-7.
• 32 McKee PA, Mattock P, Hill R. Subunit Structure of Human Fibrinogen, Soluble Fibrin, and Cross-Linked Insoluble Fibrin. Proc Nat Acad Sci USA 1970; 66: 738-44.
• 33 Folk JE, Finlayson JS. The ε-(γ-glutamyl)lysine Crosslink and the Catalytic Role of Transglutaminases. Adv Prot Chem 1977; 31: 1-133.
• 34 Kanaide H, Shainoff JR. Cross-linking of Fibrinogen and Fibrin by Fibrin-stabilizing Factor (Factor XIIIa). J Lab Clin Med 1975; 85: 574-97.
• 35 Mosesson MW, Siebenlist KR, Hainfeld JF, Wall JS. The Covalent Structure of Factor XIIIa Crosslinked Fibrinogen Fibrils. J Str Biol 1995; 115: 88-101.
• 36 Schwartz ML, Pizzo SV, Hill RL, McKee PA. Human Factor XIII from Plasma and Platelets. Molecular Weights, Subunit Structures, Proteolytic Activation, and Cross-Linking of Fibrinogen and Fibrin. J Biol Chem 1973; 248: 1395-407.
• 37 Chung SI, Lewis MS, Folk JE. Relationships of the Catalytic Properties of Human Plasma and Platelet Transglutaminases (Activated Blood Coagulation Factor XIII) to Their Subunit Structures. J Biol Chem 1974; 249: 940-50.
• 38 Cooke RD, Holbrook JJ. The Calcium-Induced Dissociation of Human Plasma Clotting Factor XIII. Biochem J 1974; 141: 79-84.
• 39 Mikuni Y, Iwanaga S, Konishi KA. Peptide Released from Plasma Fibrin Stabilzing Factor in the Conversion to the Active Enzyme by Thrombin. Biochem Biophys Res Commun 1973; 54: 1393-402.
• 40 Nakamura S, Iwanaga S, Suzuki T, Mikuni Y, Konishi K. Amino Acid Sequence of the Peptide Released from Bovine Factor XIII Following Activation by Thrombin. Biochem Biophys Res Commun 1974; 58: 250-6.
• 41 Takagi T, Doolittle RF. Amino Acid Sequence Studies on Factor XIII and the Peptide Released During Its Activation by Thrombin. Biochemistry 1974; 13: 750-6.
• 42 Lorand L, Konishi K. Activation of the Fibrin Stabilizing Factor of Plasma by Thrombin. Arch Biochem Biophys 1964; 105: 58-67.
• 43 Cooke RD. Calcium-Induced Dissociation of Human Plasma Factor XIII and the Appearance of Catalytic Activity. Biochem J 1974; 141: 683-91.
• 44 Curtis CG, Brown KL, Credo RB, Domanik A, Gray A, Stenberg P, Lorand L. Calcium-Dependent Unmasking of Active Center Cysteine During Activation of Fibrin Stabilizing Factor. Biochemistry 1974; 13: 3774-80.
• 45 Credo RB, Curtis CG, Lorand L. Ca²⁺-Related Regulatory Function of Fibrinogen. Proc Natl Acad Sci USA 1978; 75: 4234-7.
• 46 Credo RB, Curtis CG, Lorand L. Alpha-Chain Domain of Fibrinogen Controls Generation of Fibrinoligase (Coagulation Factor XIIIa). Calcium Ion Regulatory Aspects. Biochemistry 1981; 20: 3770-8.
• 47 Janus TJ, Lewis SD, Lorand L, Shafer JA. Promotion of Thrombin-Catalyzed Activation of Factor XIII by Fibrinogen. Biochemistry 1983; 22: 6269-72.
• 48 Greenberg CS, Miraglia CC, Rickles FR, Shuman MA. Cleavage of Blood Coagulation Factor XIII and Fibrinogen by Thrombin During In Vitro Clotting. J Clin Invest 1985; 75: 1463-70.
• 49 Greenberg CS, Miraglia CC. The Effect of Fibrin Polymers on Thrombin-Catalyzed Plasma Factor XIIIa Formation. Blood 1985; 66: 466-9.
• 50 Greenberg CS, Achyuthan KE, Fenton II JW. Factor XIIIa Formation Promoted by Complexing of Alpha-Thrombin, Fibrin, and Plasma Factor XIII. Blood 1987; 69: 867-71.
• 51 Greenberg CS, Achyuthan KE, Rajagopalan S, Pizzo SV. Characterization of the Fibrin Polymer Structure that Accelerates Thrombin Cleavage of Plasma Factor XIII. Arch Biochem Biophys 1988; 262: 142-8.
• 52 Lewis SD, Janus SD, Lorand L, Shafer JA. Regulation of Formation of Factor XIIIa by its Fibrin Substrates. Biochemistry 1985; 24: 6772-7.
• 53 Naski MC, Lorand L, Shafer JA. Characterization of the Kinetic Pathway for Fibrin Promotion of Alpha-Thrombin-Catalyzed Activation of Plasma Factor XIII. Biochemistry 1991; 30: 934-41.
• 54 Procyk R, Bishop PD, Kudryk B. Fibrin – Recombinant Human Factor XIII A-Subunit Association. Thromb Res 1993; 71: 127-38.
• 55 Greenberg CS, Achyuthan KE, Miraglia CC, Dobson JV. Regulation of Thrombin Cleavage of Plasma Factor XIII Bound to Fibrin. Ann NY Acad Sci 1986; 485: 140-3.
• 56 Lorand L, Credo RB, Janus TJ. Factor XIII (Fibrin-Stabilizing Factor). Meth Enzymol 1981; 80: 333-41.
• 57 Blombäck B, Procyk R, Adamson L, Hessel B. FXIII Induced Gelation of Human Fibrinogen – An Alternative Thiol Enhanced, Thrombin Independent Pathway. Thromb Res 1985; 37: 613-28.
• 58 Kazal LA, Amsel S, Miller OP, Tocantins LM. The Preparation and Some Properties of Fibrinogen Precipitated from Human Plasma by Glycine. Proc Soc Exp Biol Med 1963; 113: 989-94.
• 59 Mosesson MW, Sherry S. The Preparation and Properties of Human Fibrinogen of Relatively High Solubility. Biochemistry 1966; 5: 2829-35.
• 60 Belitser VA, Varetskaja TV, Malneva GV. Fibrinogen-Fibrin Interaction. Biochim Biophys Acta 1968; 154: 367-75.
• 61 Mosesson MW, Finlayson JS. Subfractions of Human Fibrinogen. Preparation and Analysis. J Lab Clin Med 1963; 62: 663-74.
• 62 Lorand L, Gotoh T. Fibrinoligase. The Fibrin Stabilizing Factor. Methods Enzymol 1970; 19: 770-82.
• 63 Loewy AG, Dunathan K, Kriel R, Wolfinger HL. Fibrinase I Purification of Substrate and Enzyme. J Biol Chem 1961; 236: 2625-33.
• 64 Laemmli UK. Cleavage of Structural Proteins During Assembly of the Head of Bacteriophage T4. Nature 1970; 227: 680-5.
• 65 Schwartz ML, Pizzo SV, Hill RL, McKee PA. The Subunit Structures of Human Plasma and Platelet Factor XIII (Fibrin-Stabilizing Factor). J Biol Chem 1971; 246: 5851-4.
• 66 Weber K, Osborn M. The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis. J Biol Chem 1969; 244: 4406-12.
• 67 Lorand L, Campbell-Wilkes LK, Cooperstein L. A Filter Paper Assay for Transamidating Enzymes Using Radioactive Amine Substrates. Anal Biochem 1972; 50: 623-31.
• 68 Polgár J, Hidasi V, Muszbek L. Non-proteolytic Activation of Cellular Protransglutaminase (Placenta Macrophage Factor XIII). Biochem J 1990; 267: 557-60.
• 69 Muszbek L, Polgár J, Boda Z. Platelet Factor XIII Becomes Active Without the Release of Activation Peptide During Platelet Activation. Thromb Haemost 1993; 69: 282-5.
• 70 Muszbek L, Haramura G, Polgár J. Transformation of Cellular Factor XIII into an Active Zymogen Transglutaminase in Thrombin-Stimulated Platelets. Thromb Haemost 1995; 73: 702-5.
• 71 Yee VC, Pederson LC, Bishop PD, Stenkamp RE, Teller DC. Structural Evidence that the Activation Peptide Is Not Released Upon Thrombin Cleavage of Factor XIII. Thromb Res 1995; 78: 389-97.
• 72 Lewis SD, Lorand L, Fenton JW, Shafer JA. Catalytic Competence of Human α- and γ-Thrombin in the Activation of Fibrinogen and Factor XIII. Biochemistry 1987; 26: 7597-603.
• 73 Dempfle C-E, Argiriou S, Kucher K, Müller-Peltzer H, Rübsamen K, Heene DL. Analysis of Fibrin Formation and Proteolysis During Intravenous Administration of Ancrod. Blood 2000; 96: 2793-802.
• 74 Barlow GH, Holleman WH, Lorand L. The Action of Arvin on Fibrin Stabilizing Factor (Factor XIII). Res Commun Chem Pathol Pharm 1970; 1: 39-42.
• 75 Siebenlist KR, Mosesson MW. Evidence for Intramolecular Cross-Linked Aα-γ Chain Heterodimers in Plasma Fibrinogen. Biochemistry 1996; 35: 5817-21.
• 76 Mosesson MW, Finlayson JS. Biochemical and Chromatographic Studies of Certain Activities Associated with Human Fibrinogen Preparations. J Clin Invest 1963; 42: 747-55.
• 77 Siebenlist KR, Mosesson MW, Meh DA, DiOrio JP, Albrecht RM, Olson JD. Coexisting Dysfibrinogenemia (γR275C) and Factor V Leiden Deficiency Associated with Thromboembolic Disease (Fibrinogen Cedar Rapids). Blood Coag Fibrinol 2000; 11: 293-304.
• 78 Ichinose A, Aoki N. Reversible Cross-Linking of Alpha 2-Plasmin Inhibitor to Fibrinogen by Fibrin-Stabilizing Factor. Biochim Biophys Acta 1982; 706: 158-64.
• 79 Copley AL. The Endoendothelial Fibrin Lining as the Crucial Barrier and the Role of Fibrin(Ogenin) Gels in Controlling Transcapillary Transport. Biorheology 1984; 21: 135-53.
• 80 Valenzuela R, Shainoff JR, DiBello PM, Urbanic DA, Anderson JM, Matsueda GR, Kudryk BJ. Immunoelectrophoretic and Immunohistochemical Characterizations of Fibrinogen Derivatives in Atherosclerotic Aortic Intimas and Vascular Prosthesis Pseudo-Intimas. Am J Pathol 1992; 141: 861-80.
• 81 Stathakis N, Mosesson MW, Chen AB, Galanakis DK. Cryoprecipitation of Fibrin-Fibrinogen Complexes Induced by the Cold-Insoluble Globulin of Plasma. Blood 1978; 51: 1211-22.
• 82 Stathakis N, Karamanolis D, Koukoulis G, Tsianos E. Characterization of Cryofibrinogen Isolated from Patients Plasma. Haemostasis 1981; 10: 195-201.
• 83 Shamir H, Pras M, Sohar E, Gafni J. Cryofibrinogen in Familial Mediterranean Fever. Arch Intern Med 1974; 134: 125-6.
• 84 Mosesson MW, Wautier JL, Amrani DL, Dervichian M, Cattan D. Evidence for Circulating Fibrin in Familial Mediterranean Fever. J Lab Clin Med 1982; 99: 559-67.
• 85 Babior BM, Matzner Y. The Familial Mediterranean Fever Gene – Cloned at Last. New Engl J Med 1997; 337: 1548-9.
• 86 Sohar E, Gafni J, Pras M, Heller H. Familial Mediterranean Fever. A Survey of 470 Cases and Review of the Literature. Am J Med 1967; 43: 227-53.
• 87 Moaddel M, Farrell DH, Daugherty MA, Fried MG. Interactions of Human Fibrinogens with Factor XIII: Roles of Calcium and the γ’ Peptide. Biochemistry 2000; 39: 6698-705.
• 88 Kleinman WA, Richie Jr JP. Status of Glutathione and Other Thiols and Disulfides in Human Plasma. Biochem Pharmacol 2000; 60: 19-29.