Clottability and Cross-Linking Reactivity of Fibrin(ogen) Following Differential Release of Fibrinopeptides A and B

M Furlan; T Seelich; E. A Beck

doi:10.1055/s-0038-1648078

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1976; 36(03): 582-592
DOI: 10.1055/s-0038-1648078

Original Article

Schattauer GmbH

Clottability and Cross-Linking Reactivity of Fibrin(ogen) Following Differential Release of Fibrinopeptides A and B

M Furlan

¹Central Haematology Laboratory, Inselspital and University of Bern, School of Medicine, CH-3010 Bern, Switzerland

,

T Seelich

¹Central Haematology Laboratory, Inselspital and University of Bern, School of Medicine, CH-3010 Bern, Switzerland

,

E. A Beck

¹Central Haematology Laboratory, Inselspital and University of Bern, School of Medicine, CH-3010 Bern, Switzerland

› Institutsangaben

Abstract

Summary

Human fibrinogen was treated at pH 6.0, 7.3 and 9.0 with thrombin, batroxobin (thrombinlike fraction of Bothrops atrox venom) or an extract of the venom from Ancistrodon contortrix contortrix. These three enzymes released the NH₂-terminal fibrinopeptides A and B at different rates. Thrombin-free, preactivated factor XIII (factor XIII_a) was added to incubation mixtures to stabilize resulting fibrin(ogen) aggregates. Cross-linking of γ-chains and the size of covalently linked fibrin-fibrinogen oligomers were studied in an early stage of fibrinopeptide cleavage using polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate.

Batroxobin (pH 7.3) and thrombin (pH. 6.0) preferentially released fibrinopeptide A, and resulting fibrin aggregates became rapidly insoluble. However, when fibrinopeptide B was removed with the contortrix enzyme, soluble cross-linked oligomers appeared initially. The opaque fibrin clots, produced by thrombin (pH 6.0) or contortrix procoagulant fraction (pH 7.3), were found to be devoid of α-polymers even after prolonged incubation with factor XIII_a. Our data suggest that the solubility and opacity of fibrin networks are not primarily related to the type of the cross-link (γ-γ versus α-α interactions).

Volltext

Referenzen

References
1 Bettelheim F. R. 1956; The clotting of fibrinogen. II. Fractionation of peptide material liberated. Biochimica et Biophysica Acta 19: 121.
2 Bilezikian S. B, Nossel H. L, Butler Jr V. P, Canfield R. E. 1975; Radioimmunoassay of human fibrinopeptide B and kinetics of fibrinopeptide cleavage by different enzymes. The Journal of Clinical Investigation 56: 438.
3 Collen D, Vandereycken G, De Maeyer L. 1970; Influence of hydrostatic pressure on the reversible polymerization of fibrin monomers. Nature 228: 669.
4 Cuatrecasas P, Wilchek M, Anfinsen C. B. 1968. Selective enzyme purification by affinity chromatography. Proceedings of the National Academy of Sciences. Washington: 61 636.
5 Ferry J. D, Morrison P. R. 1947; Preparation and properties of serum and plasma proteins. VIII. The conversion of human fibrinogen to fibrin under various conditions. Journal of the American Chemical Society 69: 388.
6 Gaffney P. J, Brasher M. 1974; Mode of action of ancrod as a defibrinating agent. Nature 251: 53.
7 Herzig R. H, Ratnoff O. D, Shainoff J. R. 1970; Studies on a procoagulant fraction of southern copperhead snake venom: The preferential release of fibrinopeptide B. The Journal of Laboratory and Clinical Medicine 76: 451.
8 Kanaide H, Shainoff J. R. 1975; Cross-linking of fibrinogen and fibrin by fibrin-stabilizing factor (factor XIII_a). Journal of Laboratory and Clinical Medicine 85: 574.
9 Kazama M, Langdell R. D. 1969; Purification of fibrin stabilizing factor (FSF). Federation Proceedings 28: 746.
10 Landis W. J, Waugh D. F. 1975; Interactions of bovine fibrinogen and thrombin. Early events in the development of clot structure. Archives of Biochemistry and Biophysics 168: 498.
11 Laurent T. C, Blombäck B. 1958; On the significance of the release of two different peptides from fibrinogen during clotting. Acta Chemica Scandinavica 12: 12.
12 Loewy A. G, Dunathan K, Kriel R, Wolfinger Jr H. L. 1961; Fibrinase I. Purification of substrate and enzyme. Journal of Biological Chemistry 236: 2625.
13 Ly B, Kierulf P, Jakobsen E, Gravem K. 1974; Characterization of soluble, FSF-stabilized fibrin complexes with special reference to the role of fibrinogen. Thrombosis Research 5: 21.
14 McKee P. A, Mattock P, Hill R. L. 1970; Subunit structure of human fibrinogen, soluble fibrin, and cross-linked insoluble fibrin. Proceedings of the National Academy of Sciences 66: 738.
15 McKenzie R, Pepper D. S, Kay A. B. 1974. Chromatographic and electrophoretic properties of synthetic human fibrinopeptides. Thrombosis et Diathesis haemorrhagica. Stuttgart: 32 651.
16 Mihalyi E. 1963; Cross-linking of bovine fibrinogen with formaldehyde. Acta Chemica Scandinavica 17: 277.
17 Mosesson M. W, Finlayson J. S. 1963; Subfractions of human fibrinogen. Preparation and analysis. Journal of Laboratory and Clinical Medicine 62: 663.
18 Payne J. W. 1973; Polymerization of proteins with glutaraldehyde. Soluble molecular-weight markers. Biochemical Journal 135: 867.
19 Sasaki T, Page I. H, Shainoff J. R. 1966; Stable complex of fibrinogen and fibrin. Science 152: 1069.
20 Shainoff J. R, Page I. H. 1962; Significance of cryoprofibrin in fibrinogen-fibrin conversion. Journal of Experimental Medicine 116: 687.
21 Shapiro A. L, Vinuela E, Maizel J. V. 1967; Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochemical and Biophysical Research Commmunications 28: 815.
22 Sherman L. A, Harwig S, Lee J. 1975; In vitro formation and in vivo clearance of fibrinogen: fibrin complexes. Journal of Laboratory and Clinical Medicine 86: 100.
23 Stewart G. J, Niewiarowski S. 1969; Nonenzymatic polymerization of fibrinogen by protamine sulfate. Biochimica et Biophysica Acta 194: 462.