Download PDF
Thromb Haemost 1981; 45(03): 242-246
DOI: 10.1055/s-0038-1650179
Original Article
Schattauer GmbH Stuttgart

Von Willebrand Activity of Low Molecular Weight Human Factor VIII Increases by Binding to Gold Granules

Miha Furlan
The Central Haematology Laboratory, Inselspital, Berne, Switzerland
,
Beat A Perret
The Central Haematology Laboratory, Inselspital, Berne, Switzerland
,
Eugene A Beck
The Central Haematology Laboratory, Inselspital, Berne, Switzerland
› Author Affiliations
Further Information

Publication History

Received 22 January 1981

Accepted 30 March 1981

Publication Date:
06 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

Human factor VIII/von Willebrand protein is a population of multimers which vary in size but contain apparently identical subunits. Large-molecular-weight forms possess higher ristocetin cofactor/von Willebrand activity than the native smaller oligomers. Disulfide reduction of large factor VIII multimers results in progressively decreasing molecular size and a loss of ristocetin cofactor activity. Small molecular forms of factor VIII were adsorbed onto gold granules (average diameter 20-30 nm) and thereby increased their ristocetin cofactor activity. The amount of adsorbed material and the extent of activation were dependent on the pH of the colloid suspension. The maximum recovery of von Willebrand activity was observed at pH 4.75. Aggregation of fixed human platelets by factor VIII-coated gold particles was dependent on ristocetin concentration and was not competitively inhibited by unbound low-molecular-weight factor VIII. These results suggest that the subunits of the native small factor VIII species possess potential binding affinity for platelet receptors, which is manifested following formation of large factor VIII polymers. We conclude that an optimal size of remarkably high molecular weight is required for efficient aggregation of platelets by factor VIII as occurs during the primary phase of hemostasis.

Keywords

Factor VIII/von Willebrand factor - Platelet aggregation - Gold granules
 

  • References

  • 1 Shapiro GA, Andersen JC, Pizzo SV, McKee PA. The subunit structure of normal and hemophilic factor VIII. J Clin Invest 1973; 52: 2198-2210
    CrossrefPubMedGoogle Scholar
  • 2 van MourikJ A, Bouma BN, LaBruyére WT, de GraafS, Mochtar IA. Factor VIII, a series of homologous oligomers and a complex of two proteins. Thromb Res 1974; 04: 155-164
    CrossrefPubMedGoogle Scholar
  • 3 Ekert H, Chavin SI. Changes in electrophoretic mobility of human factor VHI-related antigen: Evidence for subunit structure. Brit J. Haemat 1977; 36: 271-279
    CrossrefPubMedGoogle Scholar
  • 4 Counts RB, Paskell SL, Elgee SK. Disulfide bonds and the quaternary structure of factor Vlll/von Willebrand factor. J Clin Invest 1978; 62: 702-709
    CrossrefPubMedGoogle Scholar
  • 5 Doucet-de BruineM H M, Sixma JJ, Over J, Beeser-Visser NH. Heterogeneity of human factor VIII. II. Characterization of forms of factor VIII binding to platelets in the presence of ristocetin. J Lab Clin Med 1978; 92: 96-107
    PubMedGoogle Scholar
  • 6 Furlan M, Perret BA, Beck EA. Studies on factor VIH-related protein. III. Size distribution and carbohydrate content of human and bovine factor VIII. Biochim Biophys Acta 1979; 579: 325-333
    CrossrefPubMedGoogle Scholar
  • 7 Koutts J, Zimmerman TS. Human factor VIII antigen binding to platelet membranes: Correlation of size and function. Clin Res 1978; 26: 350
    PubMedGoogle Scholar
  • 8 Gralnick HR, Sultan Y, Coller BS. Von Willebrand’s disease. Combined qualitative and quantitative abnormalities. New Engl J Med 1977; 296: 1024-1030
    CrossrefPubMedGoogle Scholar
  • 9 Peake IR, Bloom AL. Abnormal factor VIII related antigen (FVIIIRAG) in von Willebrand’s disease (vWd): Decreased precipitation by concanavalin A. Thrombos Haemostas 1977; 37: 361-362
    PubMedGoogle Scholar
  • 10 Ruggeri ZM, Mannucci PM, Bader R, Barbui T. Factor Vlll-related properties in platelets from patients with von Willebrand’s disease. J Lab Clin Med 1978; 91: 132-140
    PubMedGoogle Scholar
  • 11 Sixma JJ, Over J, Bouma BN, Bloom AL, Peake IR. Predominance of normal low molecular weight forms of factor VIII in “variant” von Willebrand’s disease. Thromb Res 1978; 12: 929-935
    CrossrefPubMedGoogle Scholar
  • 12 Gralnick HR. Factor Vlll/von Willebrand factor protein. Galactose, a cryptic determinant of von Willebrand factor activity. J Clin Invest 1978; 62: 496-499
    CrossrefPubMedGoogle Scholar
  • 13 Sodetz JM, Paulson JC, Pizzo SV, McKee PA. Carbohydrate on human factor VHI/von Willebrand factor. Impairment of function by removal of specific galactose residues. J Biol Chem 1978; 253: 7202-7206
    PubMedGoogle Scholar
  • 14 Morisato DK, Gralnick HR. Selective binding of the factor VHI/von Willebrand factor protein to human platelets. Blood 1980; 55: 9-15
    PubMedGoogle Scholar
  • 15 Furlan M, Perret BA, Beck EA. Studies on factor VHI-related protein. IV. Interaction of galactose-specific lectins with human factor VHI/von Willebrand factor. Biochim Biophys Acta 1980; 623: 402-411
    CrossrefPubMedGoogle Scholar
  • 16 Furlan M, Jakab T, Beck EA. Effects of amidination and chemical cross-linking on human factor VIII (antihemophilic factor). Biochim Biophys Acta 1977; 494: 319-325
    CrossrefPubMedGoogle Scholar
  • 17 Horisberger M, Rosset J. Colloidal gold. A useful marker for transmission and scanning electron microscopy. J Histochem Cytochem 1977; 25: 295-305
    CrossrefPubMedGoogle Scholar
  • 18 Cooper HA, Reisner FF, Hall M, Wagner RH. Effects of thrombin treatment on preparations of factor VIII and the Ca2+-dissociated small active fragment. J Clin Invest 1975; 56: 751-760
    CrossrefPubMedGoogle Scholar
  • 19 Meyer D, Dreyfus MD, Larrieu MJ. Willebrand factor: Immunological and biological study. Path Biol 1973; 21 Suppl: 66-71
    PubMedGoogle Scholar
  • 20 Beck EA, Tranqui-Pouit L, Chapel A, Perret BA, Furlan M, Hudry-Clergeon G, Suscillon M. Studies on factor VHI-related protein. I. Ultrastructural and electrophoretic heterogeneity of human factor VHI-related protein. Biochim Biphys Acta 1979; 578: 155-163
    CrossrefPubMedGoogle Scholar
  • 21 Perret BA, Furlan M, Beck EA. Studies on factor VIII-related protein. II. Estimation of molecular size differences between factor VIII oligomers. Biochim Biophys Acta 1979; 578: 164-174
    CrossrefPubMedGoogle Scholar
  • 22 Geoghegan WD, Ackerman GA. Adsorption of horseradish peroxidase, ovomucoid and antiimmunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat antihuman immunoglobulin G on cell surfaces at the electron microscopic level: A new method, theory and application. J Histochem Cytochem 1977; 25: 1187-1200
    CrossrefPubMedGoogle Scholar
  • 23 Johnson AJ, MacDonald VE, Semar M, Fields JE, Schuck J, Lewis C, Brind J. Preparation of the major plasma fractions by solid-phase polyelectrolytes. J Lab Clin Med 1978; 92: 194-210
    PubMedGoogle Scholar
  • 24 Tan HK, Andersen JC. Human factor VIII: Morphometric analysis of purified material in solution. Science 1977; 198: 932-934
    CrossrefPubMedGoogle Scholar
  • 25 Fass DN, Knutson GJ, Bowie EJ W. Porcine Willebrand factor: A population of multimers. J Lab Clin Med 1978; 91: 307-320
    PubMedGoogle Scholar
  • 26 van MourikJ A, Bolhuis PA. Dispersity of human factor VIII – von Willebrand factor. Thromb Res 1978; 13: 15-24
    CrossrefPubMedGoogle Scholar
  • 27 Over J, Bouma BN, van MourikJ A, Sixma JJ, Vlooswijk R, Bakker-Woudenberg I. Heterogeneity of human factor VIII. I. Characterization of factor VIII present in the supernatant of cryoprecipitate. J Lab Clin Med 1978; 91: 32-46
    PubMedGoogle Scholar
  • 28 Cooper HA, Barnes DS, Wagner RH. Dithiothreitol reduction of bovine platelet aggregating factor (PAF). Blood 1975; 46: 1048
    PubMedGoogle Scholar
  • 29 Brown JE, Baugh R, Hougie C. Selective destruction of the platelet aggregation component in bovine factor VIII preparation by reduction. Thromb Res 1976; 8: 777-783
    CrossrefPubMedGoogle Scholar
  • 30 Suzuki K, Nishioka J, Hashimoto S. The influence of 2-mercaptoethanol on von Willebrand factor and bovine platelet aggregating factor. Thromb Res 1980; 17: 443-452
    CrossrefPubMedGoogle Scholar