Thromb Haemost 1967; 18(01/02): 089-100
DOI: 10.1055/s-0038-1655020
Originalarbeiten — Original Articles — Travaux Originaux
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Split Products of Fibrinogen after Prolonged Interaction with Plasmin

J.-E Niléhn
1   Coagulation Laboratory (Head: I.M. Nilsson, M.D.) and Department of Clinical Chemistry (Head: C.-B. Laurell, M.D.), University of Lund, Allmänna Sjukhuset, Malmö (Sweden)
› Author Affiliations
This investigation was supported by grants (B 67-40X-678-02; 13X-581-02A) from the Swedish Medical Research Council, from the Medical Faculty, University of Lund and the Fylgia Foundation.
Further Information

Publication History

Publication Date:
26 June 2018 (online)

Summary

End split products of fibrinogen after prolonged interaction with plasmin were separated on DEAE-cellulose. The D and E compounds measured with light ab- sorbancy constituted about 75% respectively 10% of the end split products.

The D product was separated by electrophoresis into 8 components with slight difference in charge and all of them carried the same antigenic determinants. All components were found in plasmin digested fibrinogen prepared from a pool and from single donors. Four of the ∼D components were predominant. The sedimentation coefficient of the D preparation with these 4 components was 5.3 and the fraction sedimented as an homogenous peak. Slight heterogeneity of the preparation was, however, found on determination of the diffusion coefficient. The molecular weight was calculated as 80,000. On gel filtration of the D fraction on Sephadex G 200, the D products were eluted in the beginning of the albumin peak. It is suggested that the D product which is practically homogenous on ultracentrifugation dissociates on electrophoresis. The combination of the electrophoretic, immunologic and ultracentrifugal data suggests that the fibrinogen molecule consists of a series of similar repetitive units.

The E product was homogenous in the ultracentrifuge and on electrophoresis. The sedimentation coefficient was 3.3 and the molecular weight was estimated to 40,800.

On filtration through Sephadex G 200 the E product was hardly distinguishable from the D product and was eluted together with the ablumin.

 
  • References

  • 1 Alkjaersig N, Fischer Sh. Proteolysis of fibrinogen by plasmin. Thrombos. Diathes. haemorrh. (Stuttg) 11: 284 1964;
  • 2 Archibald W. J. A demonstration of some new methods of determining molecular weights from the data of the ultracentrifuge. J. Physical and colloid chemistry 51: 1204 1947;
  • 3 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Arkiv for Kemi. 10: 415 1956;
  • 4 Caspary E. A, Kekwick R. A. Observations on the molecular weight of human fibrinogen. Biochem. J. 1954; 56 Proc. XXXV.
  • 5 Fletcher A. P. Fibrinolytic dysfibrinogenemias. Fed. Proc. 24: 822 1965;
  • 6 Garner R. L, Tillett W. Biochemical studies on the fibrinolytic activity of hemolityc streptococci. II. Nature of the reaction. J. exp. Med. 60: 255 1934;
  • 7 Hanson Z. J-B, Laurell C-B, Bachmann R. Sedimentation constants of IgG and IgD myeloma proteins compared with those of normal IgG. Acta med. scand., Suppl. 445: 89 1966;
  • 8 Hedner U, Nilsson I. M. Determination of plasminogen in human plasma by a casein method. Thrombos. Diathes. haemorrh. (Stuttg) 14: 545 1965;
  • 9 Jamieson G. A, Pert J. H. Stduies on the digestion of human fibrinogen with plasmin. Vox Sang. 08: 460 1963;
  • 10 Jamieson G. A, Gaffney P. J. Heterogeneity of fibrin polymerization inhibitor. Xlth Congress of the Internat. Soc. Haematol. 40 1966;
  • 11 Laurell C-B. Antigen antibody crossed electrophoresis. Anal. Biochem. 10: 358 1965;
  • 12 Laurell C-B. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal. Biochem. 15: 45 1966;
  • 13 Lewis J. H, Wilson J. H. Fibrinogen breakdown products. Amer. J. Physiol. 207: 1053 1964;
  • 14 Miller L, Houghton J. A. The micro-Kjeldahl determination of the nitrogen content of amino acids and proteins. J. biol. Chem. 159: 373 1945;
  • 15 Nanninga L. B. Preparation of anticoagulant split products of fibrinogen and its determination in plasma. Fed. Proc. 25: 445 1966;
  • 16 Niléhn J-E, Nilsson I. M. Demonstration of fibrinolytic split products in human serum by an immunological method in spontaneous and induced fibrinolytic states. Scand. J. Haemat. 01: 313 1964;
  • 17 Nussenzweig V, Seligmann M. Analyse, par des methodes immunochimiques de la dégradation par la plasmine du fibrinogène humain et de la fibrine, a différentes stades. Rev. Hémat. 15: 451 1960;
  • 18 Nussenzweig V, Seligmann M, Pelmont J, Grabar P. Les produits de dégradation du fibrinogène humain par la plasmine. I. Separation et propriétés physico-chimiques. Ann. Inst. Pasteur. 100: 377 1961;
  • 19 Nussenzweig V, Seligmann M, Grabar P. Les produits de dégradation du fibrinogène humain par la plasmine. II. Etude immunologique : Mise en évidence d’anticorps anti- fibrinogène natif possédant des spécificités différentes. Ann. Inst. Pasteur. 100: 490 1961;
  • 20 Salmon J. Etude immunochimique du fibrinogène et de ses dérivés. Clin. chim. Acta. 04: 767 1959;
  • 21 Scheidegger J. J. Une micro-methode de l’immunoelectrophorèse. Int. Arch. Allergy 07: 103 1955;
  • 22 SchwicJc G, Kranz Th, Schmidtberger R, StöriJco K. Zur Immunologie des Fibrinogens und seiner Spaltprodukte. Behringwerkmitteilungen 43: 213 1964;
  • 23 Seegers W. H, Nieft M. L, Vandenbeit J. M. Decomposition products of fibrinogen and fibrin. Arch. Biochem. 07: 15 1945;
  • 24 Seligmann M, Marder V. Application des techniques immunochimiques à l’étude du fibrinogène et de ses produits de degradation par la plasmine. Nouv. Rev. franc. Hémat. 05: 345 1965;
  • 25 Trautman R. Operating and comparating procedures facilitating Schlieren pattern analysis in analytical ultracentrifugation. J. Phys. Chem. 60: 1211 1956;
  • 26 Trautman R, Crampton FCh. Application of the Archibald principle for the ultracentrifugal determination of the molecular weight in urea solutions of histone fractions from calf thymus. J. Amer. chem. Soc. 81: 4036 1959;
  • 27 Triantaphyllopoulos E, Triantaphyllopoulos D. C. Variability of the anticoagulant fraction of incubated fibrinogen. Amer. J. Physiol. 208: 521 1965;
  • 28 Wadsworth C, Hanson LÅ. Comparative analysis of immune electrophoretic precipitates employing a modified immun electrophoretic technique. Int. Arch. Allergy 17: 165 1960;
  • 29 Wallen P. Studies on the purification of human plasminogen. Arkiv for Kemi. 19: 451 1962;