Download PDF
Thromb Haemost 1977; 38(02): 0524-0535
DOI: 10.1055/s-0038-1651859
Original Article
Schattauer GmbH

Fibrin and Fibrinogen Proteolysis Products: Comparison between Gel Filtration and SDS Polyacrylamide Electrophoresis Analysis

Norma Alkjaersig
1   Department of Medicine, Box 8073, Washington University School of Medicine St. Louis, Missouri 63110, U. S. A.
,
Andrew Davies   *
1   Department of Medicine, Box 8073, Washington University School of Medicine St. Louis, Missouri 63110, U. S. A.
,
Anthony Fletcher
1   Department of Medicine, Box 8073, Washington University School of Medicine St. Louis, Missouri 63110, U. S. A.
› Author Affiliations
Further Information

Publication History

Received 06 September 1976

Accepted 06 April 1977

Publication Date:
04 July 2018 (online)

    Permissions and Reprints

Summary

The proteolysis of purified human fibrinogen, stabilized and non-stabilized fibrin by plasmin were investigated by gel filtration analysis and SDS polyacrylamide electrophoresis of the reaction products. Plasmin proteolysis of fibrinogen followed the sequential steps previously reported and the two analytical methods yielded concordant results. Large molecular weight proteolysis products, of substantially greater molecular weight than native fibrinogen, were identified by gel filtration analysis following dissolution of stabilized and non-stabilized fibrin clots; with further incubation with plasmin, these proteolysis products gradually diminished in size. On the other hand, SDS polyacrylamide electrophoresis of these fibrin digests demonstrated that while non-stabilized fibrin yielded breakdown products similar in size to those obtained after proteolysis of fibrinogen, stabilized fibrin digests showed moieties of greater molecular size estimated to be of molecular weight 400,000 to 800,000. The final breakdown products of stabilized fibrin differed from those of fibrinogen and nonstabilized fibrin in that fragment D was present in the “double D” cross-linked form.

* Present address: Department of Medicine, University of Leeds, U.K.


 

  • References

  • 1 Alkjaersig N, Roy L, Fletcher A, Murphy E. 1973; Thrombosis Research. 3: 525.
    PubMedGoogle Scholar
  • 2 Budzynski A, Marder V, Shainoff J. 1974; Journal of Biological Chemistry. 249: 2294.
    PubMedGoogle Scholar
  • 3 Donati M. B, Verhaeghe R, Culasso D. E, Vermylen J. 1976; Thrombosis and Haemostasis. 36: 14.
    PubMedGoogle Scholar
  • 4 Fish W, Reynolds J, Tanford C. 1970; Journal of Biological Chemistry. 245: 5166.
    PubMedGoogle Scholar
  • 5 Fletcher A, Alkjaersig N. 1976 In: Ritchie R. (ed.) Automated Immunoanalysis. Marcel Decker, N.Y., in press.
    PubMedGoogle Scholar
  • 6 Fletcher A, Alkjaersig N, Roy L, Owens O. 1973; Proceeding of the Third Technicon Infl Symposia. Tarrytown, N.Y., Mediaid, Inc 4: 31.
    PubMedGoogle Scholar
  • 7 Gaffney P, Brasher M. 1973; Biochimica Biophysica Acta. 295: 308.
    PubMedGoogle Scholar
  • 8 Gormsen J, Feddersen C. 1974; Thrombosis Research. 5: 125.
    PubMedGoogle Scholar
  • 9 Kopec M, Teisseyre E, Dudek-Wojciechowska G, Kloczewiak M, Planklewicz V, Latallo Z. 1973; Thrombosis Research. 2: 283.
    PubMedGoogle Scholar
  • 10 Marder V, Budzynski A, Barlow G. 1975; Federation Proceedings. 34: 281.
    PubMedGoogle Scholar
  • 11 Pizzo S, Schwartz M, Hill R, McKee P. (1973a): Journal of Biological Chemistry. 248: 4574.
    PubMedGoogle Scholar
  • 12 Pizzo S, Taylor L, Schwartz M, Hill R, McKee P. (1973b): Journal of Biological Chemistry. Sachs D, Painter E. 1972; Science. 175: 781.
    PubMedGoogle Scholar
  • 13 Schwartz M, Pizzo S, Hill R, McKee P. 1971; Journal of Clinical Investigations. 50: 1506.
    PubMedGoogle Scholar
  • 14 Weber K, Osborn M. 1969; Journal of Biological Chemistry. 244: 4406.
    PubMedGoogle Scholar
  • 15 Winzor D. 1969. Molecular Biology. New York: Academic Press; 451.
    Google Scholar