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Thromb Haemost 1969; 21(03): 419-427
DOI: 10.1055/s-0038-1653556
Originalarbeiten - Original Articles - Travaux Originaux
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Bovine Platelet Proteins II. Purification of Platelet Fibrinogen

N. O Solum
1   Institute for Thrombosis Research, Rikshospitalet, Oslo 1, Norway
,
S Łopaciuk   1)
1   Institute for Thrombosis Research, Rikshospitalet, Oslo 1, Norway
› Author Affiliations
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Publication History

Publication Date:
10 June 2018 (online)

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Summary

1. Platelet fibrinogen has been purified from washed bovine platelets. The procedure was based on the methods for purification of plasma fibrinogen by fractionated precipitations and extractions with ethanol and glycine below 0°, and precipitation of proteins by dimethylformamide at 0°.

2. The platelet extract obtained by freezing and thawing of the cells, freed from insoluble material by centrifugation at 23,000 x g for 30 min, contained 0.22 ±0.003mg fibrinogen per 109 platelets. Total protein of this fraction was 0.77 ±0.08 mg per 109 platelets whereas that of the insoluble fraction was 0.79 ±0.09 mg per 109 platelets.

3. The most purified platelet fibrinogen fraction contained 91-98% of the protein in a thrombin-clottable state. The yield was approx. 20%. It showed homogeneity in analytical ultracentrifugation, in immunoelectrophoresis using an antiserum produced by immunization of rabbits against platelet extract, and in starch gel electrophoresis using a discontinuous system of Tris HCl and borate buffers offering a high resolution power towards the platelet proteins. Polyacrylamide disc electrophoresis revealed two to three faint lines behind the main fibrinogen line. At least one such line was also observed with purified plasma fibrinogen.

1) On WHO fellowship. Permanent adress: Institute of Haematology, Chocimska 5, Warsaw, Poland.


 

  • References

  • 1 Roskam J. Contribution a l’étude de la physiologie normale et pathologique du globulin (plaquette de Bizzozero). Arch. int. Physiol 20: 241 1923;
    PubMedGoogle Scholar
  • 2 Ware A. G, Fahey J. L, Seegers W. H. Platelet extracts, fibrin formation and interaction of purified prothrombin and thromboplastin. Amer. J. Physiol 154: 140 1948;
    PubMedGoogle Scholar
  • 3 Born G. V. R, Cross M. J. Effects of inorganic ions and of plasma proteins on the aggregation of blood platelets by adenosine diphosphate. J. Physiol. (Lond) 170: 397 1964;
    CrossrefPubMedGoogle Scholar
  • 4 McLean J. R, Maxwell R. E, Hertler D. Fibrinogen and adenosine diphosphate-induced aggregation of platelets. Nature (Lond) 202: 605 1964;
    CrossrefPubMedGoogle Scholar
  • 5 Solum N. O, Stormorken H. Influence of fibrinogen on the aggregation of washed human blood platelets induced by adenosine diphosphate, thrombin, collagen and adrenaline. Scand. J. clin. Lab. Invest Suppl. 84 17: 170 1965;
    PubMedGoogle Scholar
  • 6 Davey M. G, Liischer E. F. Actions of some coagulant snake venoms on blood platelets. Nature (Lond) 207: 730 1965;
    CrossrefPubMedGoogle Scholar
  • 7 Levin J. Blood coagulation and endotoxin in invertebrates. Fed. Proc 26: 1707 1967;
    PubMedGoogle Scholar
  • 8 Łopaciuk S, Solum N. O. Bovine platelet proteins. I. Soluble proteins studied by immunoelectrophoresis and polyacrylamide disc electrophoresis. Thrombos. Diathes. haemorrh. (Stuttg) 21: 409 1969;
    PubMedGoogle Scholar
  • 9 Grette K. Studies on the mechanism of thrombin-catalyzed hemostatic reactions in blood platelets. Acta physiol, scand Suppl. 195, 56 1962;
    PubMedGoogle Scholar
  • 10 Bezkorovainy A, Rafelson Jr M. E. Characterization of some proteins from normal human platelets. J. Lab. clin. Med 64: 212 1964;
    PubMedGoogle Scholar
  • 11 Laki K. The polymerization of proteins: The action of thrombin on fibrinogen. Arch. Biochem 32: 317 1951;
    CrossrefPubMedGoogle Scholar
  • 12 Davey M. G, Lüscher E. F. In: Kowalski E, Niewiarowski S. Biochemistry of blood platelets. 9 Academic Press; New York — London: 1967
    Google Scholar
  • 13 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Ark. Kemi 10: 415 1956;
    PubMedGoogle Scholar
  • 14 Blombäck B, Blombäck M, Holmberg E. A new method for fractionation of proteins. Acta chem. scand 20: 2317 1966;
    CrossrefPubMedGoogle Scholar
  • 15 Solum N. O, Łopaciuk S. Bovine platelet proteins. III. Some properties of platelet fibrinogen. Thrombos. Diathes. haemorrh. (Stuttg) 21: 428 1969;
    PubMedGoogle Scholar
  • 16 Miller G. L. Protein determinations for large numbers of samples. Analyt. Chem 31: 964 1959;
    PubMedGoogle Scholar
  • 17 Lowry O. H, Rosebrough N. J, Farr A. L, Randall R. J. Protein measurement with the Folin phenol reagent. J. biol. Chem 193: 265 1951;
    PubMedGoogle Scholar
  • 18 Smithies O. Zone electrophoresis in starch gels: Group variations in the serum proteins of normal human adults. Biochem. J 61: 629 1955;
    CrossrefPubMedGoogle Scholar
  • 19 Sasaki T, Page I. H, Shainoff J. R. Stable complex of fibrinogen and fibrin. Science 152: 1069 1966;
    CrossrefPubMedGoogle Scholar
  • 20 Mosesson M. W, Sherry S. The preparation and properties of human fibrinogen of relatively high solubility. Biochemistry 05: 2829 1966;
    CrossrefPubMedGoogle Scholar
  • 21 Mosesson M. W, Alkjaersig N, Sweet B, Sherry S. Human fibrinogen of relatively high solubility. Comparative biophysical, biochemical and biological studies with fibrinogen of lower solubility. Biochemistry 06: 3279 1967;
    PubMedGoogle Scholar
  • 22 Castaldi P. A, Caen J. Platelet fibrinogen. J. clin. Path 18: 579 1965;
    CrossrefPubMedGoogle Scholar
  • 23 Kopec M, Budzynski A, Stachurska J, Wegrzynowicks Z, Kowalski E. Studies on the mechanism of interference by fibrinogen degradation products (FDP) with the platelet function. Role of fibrinogen in the platelet atmosphere. Thrombos. Diathes. haemorrh. (Stuttg) 15: 476 1966;
    PubMedGoogle Scholar
  • 24 Nachman R. L, Marcus A. J, Zucker-Franklin D. Immunologic studies of proteins associated with subcellular fractions of normal human platelets. J. Lab. clin. Med 69: 651 1967;
    PubMedGoogle Scholar
  • 25 Holmsen H, Day H. J. Release of ß-glucuronidase from platelets induced by thrombin. Nature (Lond) 219: 760 1968;
    CrossrefPubMedGoogle Scholar