Thromb Haemost 1973; 29(02): 313-338
DOI: 10.1055/s-0038-1647774
Original Article
Schattauer GmbH

A Comparative Study of Crosslinked and Noncrosslinked Fibrin from the Major Classes of Vertebrates

Martin L. Schwartz
1   Department of Biochemistry, Duke University Medical Center, The Department of Medicine, Veterans Administration Hospital, Durham, North Carolina 27710, and the Duke University Marine Biology Laboratory, Beaufort, North Carolina
,
Salvatore V. Pizzo
1   Department of Biochemistry, Duke University Medical Center, The Department of Medicine, Veterans Administration Hospital, Durham, North Carolina 27710, and the Duke University Marine Biology Laboratory, Beaufort, North Carolina
,
J Bolling Sullivan
1   Department of Biochemistry, Duke University Medical Center, The Department of Medicine, Veterans Administration Hospital, Durham, North Carolina 27710, and the Duke University Marine Biology Laboratory, Beaufort, North Carolina
,
Robert L. Hill
1   Department of Biochemistry, Duke University Medical Center, The Department of Medicine, Veterans Administration Hospital, Durham, North Carolina 27710, and the Duke University Marine Biology Laboratory, Beaufort, North Carolina
,
Patrick A. McKee
1   Department of Biochemistry, Duke University Medical Center, The Department of Medicine, Veterans Administration Hospital, Durham, North Carolina 27710, and the Duke University Marine Biology Laboratory, Beaufort, North Carolina
› Author Affiliations
This work was supported by research grants from the National Heart and Lung Institute (HE-06400) and the National Institute of Neurological Diseases and Stroke (NS-06233), National Institutes of Health and a Clinical Investigatorship from the Veterans Administration.
Drs. Schwartz and Pizzo are predoctoral fellows, Medical Scientist Training Program, National Institute of General Medical Sciences (GM-01678).
Further Information

Publication History

Received for publication 24 August 1972

Publication Date:
24 July 2018 (online)

Summary

Crosslinked and noncrosslinked fibrin formed by clotting whole plasma in the presence and absence of calcium has been examined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The fibrin from 45 species of vertebrates with representatives from all classes except the Bradyodonti (Chimaeras) and a majority of the subclasses have been compared. Noncrosslinked fibrin from the mammals contained subunits resembling the α, β, and γ chains of human noncrosslinked fibrin. The molecular weight of the α-chains varied greatly, the molecular weight of the β-chains varied slightly, while the molecular weight of the γ-chains was apparently invariant. Nonerosslinked avian fibrins showed clearly resolved γ-chains with a molecular weight slightly smaller than human γ-chains. Avian α- and β-chains were usually not resolved. Nonerosslinked fibrin from the reptiles and amphibians contained a clearly resolved subunit with a molecular weight similar to that of human γ-chain. In crosslinked fibrin from all of the mammals, birds, reptiles, and amphibians the γ-chain was absent and a new band which corresponded to a dimer of the γ-chain was found, while high molecular weight polymers were only found in a few species. Fibrin formed from the plasma of the bony fishes was often difficult because of the problem of fibrinolysis ; however, non crosslinked fibrin from some species of bony fish had three clearly resolved subunits while the crosslinked fibrin from all of the species examined had dimers as the predominant crosslinked forms. The noncrosslinked fibrin from the cartilaginous fish had subunits in the same molecular weight range as the other vertebrate fibrins; however, the crosslinked fibrin was unique to this class because all of the fibrin subunits were involved in the crosslinking process. Dimers appeared to be a transient species, and a large number of different high molecular weight crosslinked species were formed. The crosslinked fibrin from the hagfish contained dimers and no higher molecular weight crosslinked forms. Rapid fibrinolysis complicated the interpretation of the results from all of the classes of fish.

 
  • References

  • 1 Andrew W. 1965. Comparative Haematology. Grune and Stratton; New York.:
  • 2 Archer R. K. 1970; Blood coagulation in non-human vertebrates. Symposia of the Zoological Society of London 27: 121.
  • 3 Blombäck B. 1971. Selectional trends in the structure of fibrinogen of different species. In: Schoffeniels E. (ed.), Biochemical Evolution and the Origin of Life. Elsevier Publishing Company; New York: 112.
  • 4 Blombäck B, Laurent T. C. 1958; N-terminal and light-scattering studies on fibrinogen and its transformation to fibrin. Arkiv Kemi 12: 137.
  • 5 Blombäck B, Yamashina I. 1958; The nitrogen terminal amino acids in fibrinogen and fibrin. Arkiv Kemi 12: 299.
  • 6 Cartwright T. 1970; Comparative aspects of the subunit structure of fibrinogen. Symposia of the Zoological Society of London 27: 189.
  • 7 Cartwright T, Kekwick R. G. O. 1971; A comparative study of human, cow, pig, and sheep fibrinogen. Biochimica et Biophysica Acta 236: 550.
  • 8 Caspary E. A, Kekwick R. A. 1957; Some physicochemical properties of human fibrinogen. Biochemical Journal 67: 41.
  • 9 Cohn E. J, Strong L. E, Hughes W. L, Mulford D. J, Ashworth J. N, Melin M, Taylor H. L. 1946; Preparation and properties of serum and plasma proteins. IV. A system for the separation into fractions of the protein and lipoprotein components of biological tissues and fluids. Journal of the American Chemical Society 68: 459.
  • 10 Doolittle R. F. 1963; Further studies on clotting and fibrinolysis in plasma from the smooth dogfish (Mustelis canis). British Journal of Haematology 09: 464.
  • 11 Doolittle R. F. 1965; Differences in the clotting of lamprey fibrinogen by lamprey and bovine thrombins. Biochemical Journal 94: 735.
  • 12 Doolittle R. F. 1965; a Characterization of lamprey fibrinopeptides. Biochemical Journal 94: 742.
  • 13 Doolittle R. F. 1968; Evolution of fibrinogen molecules. Thrombosis et Diathesis Haemorrhagica Supplement 39: 25.
  • 14 Doolittle R. F, Lorand L, Jacobsen A. 1963; Some comparative aspects of the fibrinogen-fibrin conversion. Biochimica et Biophysica Acta 69: 161.
  • 15 Doolittle R. F, Oncley J. L, Surgenor M. 1962; Species differences in the interaction of thrombin and fibrinogen. Journal of Biological Chemistry 237: 3123.
  • 16 Doolittle R. F, Surgenor D. M. 1962; Blood coagulation in fish. American Journal of Physiology 203: 964.
  • 17 Ende H, Meyerhoff G, Schulz G. V. 1958; Das Fibrinogenmolekül und sein Verhalten. I. Bestimmung des Molekulargewichts, des Achsenverhältnisses und der Osmotischen Virialkoeffizienten durch Messung der Sedimentation und Diffusion. Z. Naturforsch., 31 b: 713.
  • 18 Endres G. F, Scheraga H. A. 1971; Molecular weight of bovine fibrinogen by sedimentation equilibrium. Archives of Biochemistry and Biophysics 144: 519.
  • 19 Fuller G. M, Doolittle R. F. 1971; Studies of invertebrate fibrinogen. I. Purification and characterization of fibrinogen from the spiny lobster. Biochemistry 10: 1305.
  • 20 Fuller G. M, Doolittle R. F. 1971; a Studies of invertebrate fibrinogen. II. Transformation of lobster fibrinogen into fibrin. Biochemistry 10: 1311.
  • 21 Gaffney P. J, Dobos P. 1971; A structural aspect of human fibrinogen suggested by its plasmin degradation. FEBS Letters 15: 13.
  • 22 Gladner J. A. 1968. The action of thrombin on fibrinogen. In: Laki K. (ed.), Fibrinogen. Marcel Dekker; New York: 87.
  • 23 Grégoire C. 1970; Haemolymph coagulation in arthropods. Symposia of the Zoological Society of London 27: 45.
  • 24 Griffith I. P. 1972; The effect of cross-links on the mobility of proteins in dodecyl sulphate - polyacrylamide gels. Biochemical Journal 126: 553.
  • 25 Hann C. S. 1969; Peptides released during coagulation of avian and reptilian fibrinogens. Biochimica et Biophysica Acta 181: 342.
  • 26 Hawkey C. M. 1970; Fibrinolysis in animals. Symposia of the Zoological Society of London 27: 133.
  • 27 Hawkey C. M. 1970; a General Summary and Conclusions. Symposia of the Zoological Society of London 27: 217.
  • 28 Henderson K. W, Nttssbaum M. 1969; The mechanism of enhanced streptokinase-induced clot lysis following in-vitro factor XIII inactivation. British Journal of Haematology 17: 445.
  • 29 Henschen A. 1964; Peptide chains in S-sulfofibrinogen and S-sulfofibrin: isolation methods and general properties. Arkiv Kemi 22: 375.
  • 30 Hirata M, Isoda S, Kanao M, Shimizur H, Inoue S. 1970; Studies on anesthetics for fish. Bulletin of the Japanese Society of Scientific Fisheries 36: 1127.
  • 31 Jensen D. 1966; The hagfish. Scientific American 214: 82.
  • 32 Lazar G. 1970; Species specificity of fibrinogen. Acta Physiologica of the Academy of Sciences of Hungary 38: 1.
  • 33 Lender T, Bouchard-Madrelle C. 1964; Etude experimental de la régénération du complex neural de Ciona intestinalis. Bulletin of the Zoological Society of France 89: 546.
  • 34 Lorand L, Downey J, Gotoh T, Jacobsen A, Tokura S. 1968; The transpeptidase system which crosslinks fibrin by γ-glutamyl-e-lysine bonds. Biochemical and Biophysical Research Communications 31: 222.
  • 35 Matacic S, Loewy A. G. 1968; The identification of isopeptide crosslinks in insoluble fibrin. Biochemical and Biophysical Research Communications 30: 356.
  • 36 McKee P. A, Mattock P, Hill R. L. 1970; Subunit structure of human fibrinogen, soluble fibrin, and crosslinked insoluble fibrin. Proceedings of the National Academy of Sciences, U.S.A 66: 738.
  • 37 McKee P. A, Rogers L. A, Marler E, Hill R. L. 1966; The subunit polypeptides of human fibrinogen. Archives of Biochemistry and Biophysics 116: 271.
  • 38 Mills D, Karpatkin K. Heterogeneity of human fibrinogen: possible relation to proteolysis by thrombin and plasmin as studied by SDS-polyacrylamide gel electrophoresis. Biochemical and Biophysical Research Communications. 40. 206.
  • 39 Needham A. E. 1970; Haemostatic mechanisms in the invertebrata. Symposia of the Zoological Society of London 27: 19.
  • 40 Niewiarowski S, Latallo Z. 1959; Comparative studies of the fibrinolytic system of sera of various vertebrates. Thrombosis et Diathesis Haemorrhagica 03: 404.
  • 41 Okijde M, Iwanaga S. 1971; Carboxyl-terminal residues of mammalian fibrinogen and fibrin. Biochimica et Biophysica Acta 251: 185.
  • 42 Pirkle H, Henschen A. 1968; N-terminal sequences of bovine fibrinogen. Biochemistry 07: 1362.
  • 43 Pirkle H, Henschen A, Potapous A. 1969; Interspecies constancy of N-terminal amino-acid sequences in the γ-chain of fibrinogen. Nature 223: 400.
  • 44 Pisano J. J, Finlayson J. S, Peyton M. P. 1969; Chemical and enzymic detection of protein crosslinks. Measurement of e - ( γ - glutamyl)ly sine in fibrin polymerized by factor XIII. Biochemistry 08: 871.
  • 45 Pizzo S. V, Schwartz M. L, Hill R. L, McKee P. A. 1972; The effect of plasmin on the subunit structure of human fibrinogen. The Journal of Biological Chemistry 247: 636.
  • 46 Pizzo S. V, Schwartz M. L, Hill R. L, McKee P. A. 1972; a Comparison of the subunit structures of plasmin digested fibrinogen and fibrin. Federation Proceedings 31: 217.
  • 47 Pizzo S. V, Schwartz M. L, Hill R. L, McKee P. A. 1972 b The effect of plasmin on the subunit structure of human fibrin. (Manuscript submitted for publication).
  • 48 Rothschild, Lord 1965. A Classification of Living Animals. Robert MacLehose and Co. Ltd., The University Press; Glasgow.:
  • 49 Schwartz M. L, Pizzo S. V, Hill R. L, McKee P. A. 1971; The effect of fibrin stabilizing factor on the subunit structure of human fibrin. Journal of Clinical Investigation 50: 1506.
  • 50 Schwartz M. L, Pizzo S. V, Hill R. L, McKee P. A. 1972 Human factor XIII from plasma and platelets. Molecular weights, subunit structures, proteolytic activation, and crosslinking of fibrinogen and fibrin. (Manuscript submitted foi publication).
  • 51 Shtjlman S. 1953; The size and shape of bovine fibrinogen. Studies of sedimentation, diffusion, and viscosity. Journal of the American Chemical Society 75: 5846.
  • 52 Solum N. O. 1970; Coagulation in Limulus — Some properties of the clottable protein of Limulus polyphemus blood cells. Symposia of the Zoological Society of London 27: 207.
  • 53 Weber K, Osborn M. 1969; The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. The Journal of Biological Chemistry 244: 4406.
  • 54 Weiner A. M, Platt T, Weber K. 1972; Amino-terminal sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. The Journal of Biological Chemistry 247: 3242.