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Thromb Haemost 1980; 44(03): 119-124
DOI: 10.1055/s-0038-1650100
Original Article
Schattauer GmbH Stuttgart

Fibrin Assembly: A Comparison of Electron Microscopic and Light Scattering Results

Roy Hantgan
*   The Department of Biochemistry, School of Medicine, University of North Carolina, Chapel Hill, NC, U.S.A.
,
Walter Fowler
**   The Departments of Pharmacology and Anatomy, Duke University Medical Center, Durham, NC, U.S.A.
,
Harold Erickson
**   The Departments of Pharmacology and Anatomy, Duke University Medical Center, Durham, NC, U.S.A.
,
Jan Hermans
*   The Department of Biochemistry, School of Medicine, University of North Carolina, Chapel Hill, NC, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 04 August 1980

Accepted 20 September 1980

Publication Date:
13 July 2018 (online)

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Summary

To test the validity of a proposed two step fibrin assembly mechanism and directly visualize the molecular species present at various stages of fibrin formation, we have carried out an electron microscopic investigation. Assembly conditions duplicated those of a recent light scattering study and specimens were prepared at different time points with the use of a negative staining technique recently employed to visualize the trinodular structure of fibrinogen. Under near-physiological buffer conditions, protofibrils structurally similar to those postulated by Ferry have been found at early stages of fibrin assembly. In parallel with the light scattering results, a dramatic increase in fiber diameter was found in specimens prepared during the postulated lateral association stage of gelation. Light scattering and electron microscopic results both showed that high ionic strength reduces the rate and extent of fiber formation. Reptilase cleavage is shown to result in typical cross striated fibrin.

Key words

Fibrinogen - Fibrin assembly - Electron microscopy - Light scattering
 

  • References

  • 1 Scheraga HA, Laskowski Jr M. The fibrinogen-fibrin conversion. Adv Prot Chem 1957; 12: 1-131
    PubMedGoogle Scholar
  • 2 Laki K, Gladner JA. Chemistry and physiology of the fibrinogenfibrin transition. Physiol Rev 1964; 44: 127-160
    CrossrefPubMedGoogle Scholar
  • 3 Doolittle RF. Structural aspects of the fibrinogen to fibrin conversion. Adv Prot Chem 1973; 27: 1-109
    PubMedGoogle Scholar
  • 4 Hantgan RR, Hermans J. Assembly of fibrin. A light scattering study. J Biol Chem 1979; 254: 11272-11281
    PubMedGoogle Scholar
  • 5 Ferry JD. The mechanism of polymerization of fibrinogen. Proc Nat Acad of Sci, USA 1952; 38: 566-569
    CrossrefPubMedGoogle Scholar
  • 6 Krakow W, Endres GF, Siegel BM, Scheraga HA. An electron microscopic investigation of the polymerization of bovine fibrin monomer. J Mol Biol 1972; 17: 95-103
    PubMedGoogle Scholar
  • 7 Blömback B, Hessel B, Hogg D, Therkildsen L. A two-step fibrinogen-fibrin transition in blood coagulation. Nature 1978; 275: 501-505
    CrossrefPubMedGoogle Scholar
  • 8 Martinelli RA, Scheraga HA. Steady-state kinetic study of the bovine thrombin-fibrinogen interaction. Biochemistry 1980; 19: 2343-2350
    CrossrefPubMedGoogle Scholar
  • 9 Fowler WE, Erickson HP. The trinodular structure of fibrinogenconformation by both shadowing and negative stain electron microscopy. J Mol Biol 1979; 134: 241-249
    CrossrefPubMedGoogle Scholar
  • 10 Siegel BM, Mernan JP, Scheraga HA. The configuration of native and partially polymerized fibrinogen. Biochim Biophys Acta 1953; 11: 329-336
    CrossrefPubMedGoogle Scholar
  • 11 Bang NV. A molecular model of fibrin based on electron microscopy of fibrin polymerization. Thromb Diath Haemorrh, Suppl 1964; 13: 73-80
    PubMedGoogle Scholar
  • 12 Pouit L, Marcille G, Suscillon M, Hollard D. Zude en microscopie électronique de différentes étapes de la fibrinoformation. Thromb Diath Haemorrh 1972; 27: 559-572
    PubMedGoogle Scholar
  • 13 Tranqui L, Prandini MH, Suscillon M. Structure of fibrinogen and its derivatives; an electron microscopic study. Thromb Haemostas 1979; 41: 655-661
    PubMedGoogle Scholar
  • 14 Hantgan RR, Hermans J, Fowler W, Erickson H. Fibrin formation as a biological assembly process. Biophys J 1980 to the published in October issue
    PubMedGoogle Scholar
  • 15 Wrigley NC. The lattice spacing of crystalline catalase as an internal standard of length in electron microscopy. J Ultrastruct Res 1968; 24: 454-464
    CrossrefPubMedGoogle Scholar
  • 16 Christiansen G, Griffith J. Salt and divalent cations affect the flexible nature of the natural beaded chromatin structure. Nucleic Acids Res 1977; 4: 1837
    CrossrefPubMedGoogle Scholar
  • 17 Shen LL, Hermans J, McDonagh J, McDonagh RP. Role of fibrinopeptide B release: comparison of fibrins produced by thrombin and ancrod. Am J Physiol 1977; 6: H629-H633
    PubMedGoogle Scholar
  • 18 Lorand L. Fibrin Clots. Nature 1950; 166: 694
    CrossrefPubMedGoogle Scholar
  • 19 Gollwitzer R, Karges HE, Horman H, Kuhn K. Electron microscopic investigations on soluble and insoluble bovine fibrin. Biochim Biophys Acta 1970; 207: 445-455
    CrossrefPubMedGoogle Scholar
  • 20 Lorand LT, Downey J, Gotoh T, Jacobsen A, Tokura S. The transpeptidase system which crosslinks fibrin by γ-glutamyl-ε-lysine bonds. Biochem Biophys Res Comm 1968; 31: 222-229
    CrossrefPubMedGoogle Scholar
  • 21 Slayter H. High-resolution metal replication of macromolecules. Ultramicroscopy 1976; 1: 341-357
    CrossrefPubMedGoogle Scholar
  • 22 Fowler WE, Hantgan RR, Hermans J, Erickson HP. (manuscript in preparation, 1980)
    PubMed
  • 23 Manyakov VF, Varetskaja TV, Belitser VA. Electron microscopic investigations on fibrin self assembly. Mol Biol (Mosk) 1977; 11: 908-913
    PubMedGoogle Scholar
  • 24 Hawn CV Z, Porter KR. The fine structure of clots formed from purified bovine fibrinogen and thrombin. A study with the electron microscope. J Exp Med 1947; 86: 285-292
    CrossrefPubMedGoogle Scholar
  • 25 Stryer L, Cohen C, Langridge R. Axial period of fibrinogen and fibrin. Nature 1963; 197: 793-794
    CrossrefPubMedGoogle Scholar
  • 26 Kay D, Cuddigan BJ. The fine structure of fibrin. Bri J Haematol 1967; 13: 341-347
    PubMedGoogle Scholar
  • 27 Flory PJ. Principles of Polymer Chemistry. 1953: 318-323 Cornell Univ. Press, Ithaca, NY:
    Google Scholar
  • 28 Carr Jr ME, Shen LL, Hermans J. A physical standard of fibrinogen: measurement of the elastic modulus of dilute fibrin gels with a new elastometer. Analyt Biochem 1976; 72: 202-211
    CrossrefPubMedGoogle Scholar