Download PDF
Thromb Haemost 2002; 87(04): 651-658
DOI: 10.1055/s-0037-1613062
Review Article
Schattauer GmbH

Fibrinogen Assembly and Crosslinking on a Fibrin Fragment E Template

Michael W. Mosesson
1   The Blood Research Institute of the Blood Center of Southeastern Wisconsin, Milwaukee, WI
,
Kevin R. Siebenlist
2   Department of Biomedical Science, College of Health Sciences, Marquette University, Milwaukee, WI
,
Irene Hernandez
1   The Blood Research Institute of the Blood Center of Southeastern Wisconsin, Milwaukee, WI
,
Joseph S. Wall
3   Biology Department, Brookhaven National Laboratory, Upton, NY, USA
,
James F. Hainfeld
3   Biology Department, Brookhaven National Laboratory, Upton, NY, USA
› Author Affiliations
Further Information

Publication History

Received 28 September 2001

Accepted after revision 14 December 2001

Publication Date:
08 December 2017 (online)

    Permissions and Reprints

Summary

There is an ongoing controversy concerning whether crosslinked γ chains in fibrin are oriented “transversely” between fibril strands or “end-to-end” along fibril strands. From the latter viewpoint, Veklich et al. [Proc Natl Acad Sci (USA) 95: 1438, 1998] observed that fibrinogen fibrils that had been assembled on a fibrin fragment E template, crosslinked with factor XIIIa, and then dissociated in acetic acid solution, were aligned end-to-end. This led to the conclusion that crosslinked γ chains in fibrin under physiological conditions were also aligned endto-end. To assess its validity we studied the assembly and organization of fibrinogen molecules on a des AB-fibrin fragment E (E-des AB) or a des A-fibrin fragment E (E-des A) template.

We evaluated the roles of E polymerization sites EA and EB, and D association sites γXL, Da, Db, βC, and αC in this process. EA:Da interactions caused fibrinogen: E “DED” complexes to form, and markedly enhanced the γ chain crosslinking rates of fibrinogen or des αC-fibrinogen. Fibrinogen crosslinking without added fibrin E was slower, and that of des αC-fibrinogen was still slower. These events showed that although αC domains promote fibrinogen fibril assembly and crosslinking, they contribute little to increasing the EA:Da-dependent crosslinking rate. Electron microscopic (STEM) images of E-des AB and fibrinogen plus factor XIIIa showed single-, double-, and multistranded fibrils with interstrand DED complexes aligned side-to-side. This alignment was due to βC:βC contacts resulting from D subdomain rearrangements initiated by the EB:Db interactions, and also occurred in mixtures of des αC-fibrinogen with E-des AB.

In contrast, a mixture of fibrinogen and E-des A plus XIIIa revealed double-stranded fibrils with interstrand DED complexes in a halfstaggered arrangement, an alignment that we attribute to crosslinking of γXL sites bridging between fibrils strands. These and other features of E-des A-based fibrinogen fibrils, including interstrand γ chain bridges and early and extensive lateral fibril strand associations concomitant with accelerated γ chain crosslinking, indicate that crosslinking of fibrin fibril strands takes place preferentially on transversely positioned γ chains.

Keywords

Factor XIII - fibrinogen - fibrin - electron microscopy
 

  • References

  • 1 Scheraga HA, Laskowski MJr. The fibrinogen-fibrin conversion. Adv Prot Chem 1957; 12: 1-131.
    PubMedGoogle Scholar
  • 2 Blombäck B. Studies on the action of thrombotic enzymes on bovine fibrinogen as measured by N-terminal analysis. Arkiv Kemi 1958; 12: 321-35.
    PubMedGoogle Scholar
  • 3 Blombäck B, Hessel B, Hogg D, Therkildsen L. A two-step fibrinogenfibrin transition in blood coagulation. Nature 1978; 275: 501-5.
    CrossrefPubMedGoogle Scholar
  • 4 Shimizu A, Nagel GM, Doolittle RF. Photoaffinity labeling of the primary fibrin polymerization site: Isolation of a CNBr fragment corresponding to γ 337-379. Proc Natl Acad Sci USA 1992; 89: 2888-92.
    CrossrefPubMedGoogle Scholar
  • 5 Pratt KP, Côté HCF, Chung DW, Stenkamp RE, Davie EW. The primary fibrin polymerization pocket: three-dimensional structure of a 30-kDa C-terminal γ chain fragment complexed with the peptide gly-pro-arg-pro. Proc Natl Acad Sci USA 1997; 94: 7176-81.
    CrossrefPubMedGoogle Scholar
  • 6 Everse SJ, Spraggon G, Veerapandian L, Riley M, Doolittle RF. Crystal structure of fragment double-D from human fibrin with two different bound ligands. Biochemistry 1998; 37: 8637-42.
    CrossrefPubMedGoogle Scholar
  • 7 Ferry JD. The mechanism of polymerization of fibrinogen. Proc Natl Acad Sci USA 1952; 38: 566-9.
    CrossrefPubMedGoogle Scholar
  • 8 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.
    CrossrefPubMedGoogle Scholar
  • 9 Fowler WE, Hantgan RR, Hermans J. et al. Structure of the fibrin protofibril. Proc Natl Acad Sci USA 1981; 78: 4872-6.
    CrossrefPubMedGoogle Scholar
  • 10 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.
    CrossrefPubMedGoogle Scholar
  • 11 Medved LV, Litvinovich SV, Ugarova TP, Lukinova NI, Kilikhevich VN, Ardemasova ZA. Localization of a fibrin polymerization site complimentary to Gly-His-Arg sequence. FEBS Letters 1993; 320: 239-42.
    CrossrefPubMedGoogle Scholar
  • 12 Yang Z, Mochalkin I, Doolittle RF. A model of fibrin formation based on crystal structures of fibrinogen and fibrin fragments complexed with synthetic peptides. Proc Natl Acad Sci USA 2000; 97: 14156-61.
    CrossrefPubMedGoogle Scholar
  • 13 Weisel JW, Medved L. The structure and function of the αC domain of fibrinogen. Ann NY Acad Sci 2001; 936: 312-27.
    PubMedGoogle Scholar
  • 14 Mosesson MW, Finlayson JS, Umfleet RA, Galanakis DK. Human fibrinogen heterogeneities. I. Structural and related studies of plasma fibrinogens which are high solubility catabolic intermediates. J Biol Chem 1972; 247: 5210-9.
    PubMedGoogle Scholar
  • 15 Mosesson MW, Galanakis DK, Finlayson JS. Comparison of human plasma fibrinogen subfractions and early plasmic fibrinogen derivatives. J Biol Chem 1974; 249: 4656-64.
    PubMedGoogle Scholar
  • 16 Mosesson MW, Sherry S. The preparation and properties of human fibrinogen of relatively high solubility. Biochemistry 1966; 05: 2829-35.
    CrossrefPubMedGoogle Scholar
  • 17 Mosesson MW. Fibrinogen Heterogeneity. Ann NY Acad Sci 1983; 408: 97-113.
    CrossrefPubMedGoogle Scholar
  • 18 Hasegawa N, Sasaki S. Location of the binding site “b” for lateral polymerization of fibrin. Thromb Res 1990; 57: 183-95.
    CrossrefPubMedGoogle Scholar
  • 19 Medved LV, Gorkun OV, Manyakov VF, Belitser VA. The role of fibrinogen αC-domains in the fibrin assembly process. FEBS Letter 1985; 181: 109-12.
    CrossrefPubMedGoogle Scholar
  • 20 Mosesson MW, Siebenlist KR, Hainfeld JF, Wall JS. The covalent structure of factor XIIIa crosslinked fibrinogen fibrils. J Struct Biol 1995; 115: 88-101.
    CrossrefPubMedGoogle Scholar
  • 21 Samokhin GP, Lorand L. Contact with the N termini in the central E domain enhances the reactivities of the distal D domains of fibrin to factor XIIIa. J Biol Chem 1995; 270: 21827-32.
    CrossrefPubMedGoogle Scholar
  • 22 Kanaide H, Shainoff JR. Cross-linking of fibrinogen and fibrin by fibrinstabilizing factor (factor XIIIa). J Lab Clin Med 1975; 85: 574-97.
    PubMedGoogle Scholar
  • 23 Mosesson MW, Siebenlist KR, Meh DA. The structure and biological features of fibrinogen and fibrin. Ann NY Acad Sci 2001; 936: 11-30.
    PubMedGoogle Scholar
  • 24 Veklich YI, Ang EK, Lorand L, Weisel JW. The complementary aggregation sites of fibrin investigated through examination of polymers of fibrinogen with fragment E. Proc Natl Acad Sci USA 1998; 95: 1438-42.
    CrossrefPubMedGoogle Scholar
  • 25 Lorand L, Parameswaren KN, Murthy SNP. A double-headed Gly-Pro-Arg-Pro ligand mimics the functions of the E domain of fibrin for promoting the end-to-end crosslinking of γ chains by factor XIIIα . Proc Natl Acad Sci USA 1998; 95: 537-41.
    CrossrefPubMedGoogle Scholar
  • 26 Siebenlist KR, Meh DA, Mosesson MW. Plasma factor XIII binds specifically to fibrinogen molecules containing γ’ chains. Biochemistry 1996; 35: 10448-53.
    CrossrefPubMedGoogle Scholar
  • 27 Henschen A, Lottspeich F, Kehl M, Southan C, Lucas J. Structure-functionevolution relationship in fibrinogen. In: Fibrinogen. Recent biochemical and medical aspects. Henschen A, Graeff H, Lottspeich F. Berlin: W de Gruyter; 1982: 67-82.
    Google Scholar
  • 28 Veklich YI, Gorkun OV, Medved LV, Niewenhuizen W, Weisel JW. Carboxyl-terminal portions of the a chains of fibrinogen and fibrin. J Biol Chem 1993; 268: 13577-85.
    PubMedGoogle Scholar
  • 29 Lorand L, Gotoh T. Fibrinoligase. The fibrin stabilizing factor. Methods Enzymol 1970; 19: 770-82.
    PubMedGoogle Scholar
  • 30 Loewy AG, Dunathan K, Kriel R, Wolfinger HL. Fibrinase I. Purification of substrate and enzyme. J Biol Chem 1961; 236: 2625-33.
    PubMedGoogle Scholar
  • 31 Malinconico SM, Katz JB, Budzynski AZ. Hementin: anticoagulant protease from the salivary gland of the leech Haementeria ghilianii. J Lab Clin Med 1984; 103: 44-58.
    PubMedGoogle Scholar
  • 32 Marder VJ, Budzynski AZ, James HL. High molecular weight derivatives of human fibrinogen produced by plasmin: (III) Their NH2-terminal amino acids and comparison with the NH2-terminal disulfide knot. J Biol Chem 1972; 247: 4775-81.
    PubMedGoogle Scholar
  • 33 Townsend RR, Hilliker E, Li YT, Laine RA, Bell WR, Lee YC. Carbohydrate structure of human fibrinogen. J Biol Chem 1982; 257: 9704.
    PubMedGoogle Scholar
  • 34 Kirschbaum NE, Budzynski AZ. A unique proteolytic fragment of human fibrinogen containing the Aα COOH-terminal domain of the native molecule. J Biol Chem 1990; 265: 13669-76.
    PubMedGoogle Scholar
  • 35 Mosesson MW, Hainfeld JF, Haschemeyer RH, Wall JS. Identification and mass analysis of human fibrinogen molecules and their domains by scanning transmission electron microscopy. J Mol Biol 1981; 153: 695-718.
    CrossrefPubMedGoogle Scholar
  • 36 Hainfeld JF, Wall JS, Desmond EJ. A small computer system for micrograph analysis. Ultramicroscopy 1982; 08: 263-70.
    CrossrefPubMedGoogle Scholar
  • 37 Wall JS, Hainfeld JF. Mass mapping with the scanning transmission microscope. Ann Rev Biophys Biophys Chem 1986; 15: 355-76.
    CrossrefPubMedGoogle Scholar
  • 38 Mosesson MW, Siebenlist KR, Hainfeld JF. et al. The relationship between the fibrinogen D domain self-association/crosslinking site (γXL) and the fibrinogen Dusart abnormality (Aα R554C-albumin). J Clin Invest 1996; 97: 2342-50.
    CrossrefPubMedGoogle Scholar
  • 39 Mosesson MW, Siebenlist KR, Meh DA, Wall JS, Hainfeld JF. The location of the carboxy-terminal region of γ chains in fibrinogen and fibrin D domains. Proc Natl Acad Sci USA 1998; 95: 10511-6.
    CrossrefPubMedGoogle Scholar
  • 40 Mosesson MW, Fass DN, Lollar P. et al. Structural model of porcine factor VIII and factor VIIIa molecules based on scanning transmission electron microscope (STEM) images and STEM mass analysis. J Clin Invest 1990; 85: 1983-90.
    CrossrefPubMedGoogle Scholar
  • 41 Siebenlist KR, Meh DA, Wall JS, Hainfeld JF, Mosesson MW. Orientation of the carboxy-terminal regions of fibrin γ chain dimers determined from the crosslinked products formed in mixtures of fibrin, fragment D, and factor XIIIa. Thromb Haemost 1995; 74: 1113-9.
    Article in Thieme ConnectPubMedGoogle Scholar