Subscribe to RSS
DOI: 10.1055/s-0038-1651859
Fibrin and Fibrinogen Proteolysis Products: Comparison between Gel Filtration and SDS Polyacrylamide Electrophoresis Analysis
Publication History
Received
06 September 1976
Accepted
06 April 1977
Publication Date:
04 July 2018 (online)
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.
- 2 Budzynski A, Marder V, Shainoff J. 1974; Journal of Biological Chemistry. 249: 2294.
- 3 Donati M. B, Verhaeghe R, Culasso D. E, Vermylen J. 1976; Thrombosis and Haemostasis. 36: 14.
- 4 Fish W, Reynolds J, Tanford C. 1970; Journal of Biological Chemistry. 245: 5166.
- 5 Fletcher A, Alkjaersig N. 1976 In: Ritchie R. (ed.) Automated Immunoanalysis. Marcel Decker, N.Y., in press.
- 6 Fletcher A, Alkjaersig N, Roy L, Owens O. 1973; Proceeding of the Third Technicon Infl Symposia. Tarrytown, N.Y., Mediaid, Inc 4: 31.
- 7 Gaffney P, Brasher M. 1973; Biochimica Biophysica Acta. 295: 308.
- 8 Gormsen J, Feddersen C. 1974; Thrombosis Research. 5: 125.
- 9 Kopec M, Teisseyre E, Dudek-Wojciechowska G, Kloczewiak M, Planklewicz V, Latallo Z. 1973; Thrombosis Research. 2: 283.
- 10 Marder V, Budzynski A, Barlow G. 1975; Federation Proceedings. 34: 281.
- 11 Pizzo S, Schwartz M, Hill R, McKee P. (1973a): Journal of Biological Chemistry. 248: 4574.
- 12 Pizzo S, Taylor L, Schwartz M, Hill R, McKee P. (1973b): Journal of Biological Chemistry. Sachs D, Painter E. 1972; Science. 175: 781.
- 13 Schwartz M, Pizzo S, Hill R, McKee P. 1971; Journal of Clinical Investigations. 50: 1506.
- 14 Weber K, Osborn M. 1969; Journal of Biological Chemistry. 244: 4406.
- 15 Winzor D. 1969. Molecular Biology. New York: Academic Press; 451.