The Interaction of the Factor Vlll/von Willebrand Factor Complex with Hematin

David Green; Fiona H Furby; Michael C Berndt

doi:10.1055/s-0038-1661666

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1986; 56(03): 277-282
DOI: 10.1055/s-0038-1661666

Original Article

Schattauer GmbH Stuttgart

The Interaction of the Factor Vlll/von Willebrand Factor Complex with Hematin

Authors

David Green

The Department of Medicine, University of Sydney, Westmead Centre, Sydney, Australia, and the Atherosclerosis Program, Department of Medicine, Northwestern University Medical School and the Rehabilitation Institute, Chicago, Illinois, USA
Fiona H Furby

The Department of Medicine, University of Sydney, Westmead Centre, Sydney, Australia, and the Atherosclerosis Program, Department of Medicine, Northwestern University Medical School and the Rehabilitation Institute, Chicago, Illinois, USA
Michael C Berndt

The Department of Medicine, University of Sydney, Westmead Centre, Sydney, Australia, and the Atherosclerosis Program, Department of Medicine, Northwestern University Medical School and the Rehabilitation Institute, Chicago, Illinois, USA

Abstract

PDF Download

Summary

Intravenous infusion of hematin, used in the treatment of acute porphyria, induces a decline in the plasma factor VIH/von Willebrand factor complex (VIII/vWF) and thrombocytopenia. We investigated this problem by studying the interaction between hematin, purified VIII/vWF, and platelets in vitro. Hematin was labeled with either ⁵⁹Fe or ³H and characterized by gel chromatography. Hematin self-aggregated, forming a complex with an average molecular weight of approximately 10,000 daltons. When incubated with VIII/vWF for 30 min at 37° C and applied to Sepharose CL-4B, the hematin eluted with the VIII/vWF in the void volume. Hematin inhibited the dissociation of factor VIII antigen (VIII :Ag) from the von Willebrand antigen (vWF:Ag) in 0.25 M CaCl₂, and reversed the aggregation of VIII:Ag induced by 0.1 M 6-aminocaproic acid. Both hematin and the hematin-VIII/vWF complex bound to washed normal platelets and to platelets from a patient with Bemard-Soulier syndrome. Thromb-asthenic platelets were not aggregatable by hematin, and bound significantly less hematin-VIII/vWF than normal platelets suggesting that hematin-induced platelet activation was required for binding. Likewise, binding was inhibited by PGE₁ which also prevented aggregation. We conclude that hematin forms complexes with VIII/vWF, alters the functional activity and dissociation of this compound, and participates in the binding of VIII/vWF to platelets.

Keywords

Factor VIII/vWF - Hematin - Platelets - Porphyria

PDF (1261 kb)

References

References
1 Bonkowsky HL, Tschudy DP, Collins A, Bossenmaier I, Cardinal R, Watson CJ. Repression of the overproduction of porphyrin precursors in acute intermittent porphyria by intravenous infusion of hematin. Proc Nat Acad Sci USA 1971; 68: 2725-2729
2 Morris DL, Dudley MD, Pearson RD. Coagulopathy associated with hematin treatment for acute intermittent porphyria. Ann Intern Med 1981; 95: 700-702
3 Glueck R, Green D, Cohen I, Ts’ao C. Hematin: unique effects on hemostasis. Blood 1983; 61: 243-249
4 Green D, Reynolds N, Klein J, Kohl H, Ts’ao C. The inactivation of hemostatic factors by hematin. J Lab Clin Med 1983; 102: 361-369
5 Neely SM, Gardner DV, Reynolds N, Green D, Ts’ao C. Mechanism and characteristics of platelet activation by hematin. Br J Haemat 1984; 58: 305-316
6 Newman J, Johnson AJ, Karpatkin MH, Palskin S. Methods for the production of clinically effective intermediate and high purity concentrates. Br J Haematol 1971; 21: 1-20
7 Booth WJ, Furby FH, Berndt MC, Castaldi PA. Factor VIII/von Willebrand factor has potent lectin activity. Biochem Biophys Res Comm 1984; 118: 495-501
8 Bothwell TH, Charlton RW, Cook JD, Finch CA. Iron metabolism in man. Blackwell Sci Publ; Oxford: 1979: 401-438
9 Fischer H. Organic synthesis collective volume. John Wiley & Sons; New York: 1955. Vol. 3 442
10 Schwartz S, Berg MH, Bossenmaier I, Dinsmore H. Determinations of porphyrins in biological materials. In: Methods of Biochemical Analysis Glick D. (ed) Interscience Publishers; New York: 1960. Vol. 8 221
11 Goldstein GW, Hammaker L, Schmid R. The catabolism of Heinz bodies: an experimental model demonstrating conversion to nonbilirubin catabolites. Blood 1968; 31: 388-395
12 Farrell GC, Gollan JL, Schmid R. Efflux of bilirubin into plasma following hepatic degradation of exogenous heme. Pro Soc Exp Biol Med 1980; 163: 504-509
13 Harrison RL, McKee PA. Comparison of thrombin and ristocetin in the interaction between von Willebrand factor and platelets. Blood 1983; 62: 346-353
14 Owen WC, Wagner RH. Antihemophilic factor: separation of an active fragment following dissociation by salts or detergents. Throm-bos Diathes Haemorrh 1972; 27: 502-515
15 Austen DE G. Factor VIII of small moplecular weight and its aggregation. Br J Haematol 1974; 27: 89-100
16 Furby FH, Berndt MC, Castaldi PA, Koutts J. Characterization of calcium dependent binding of endogenous factor VIII/von Willebrand factor to surface activated platelets. Thromb Res 1984; 35: 501-511
17 Berndt MC, Gregory C, Chong BH, Zola H, Castaldi PA. Additional glycoprotein defects in Bemard-Soulier’s Syndrome: confirmation of genetic basis by parental analysis. Blood 1983; 62: 800-807
18 Reimers HJ, Packham MA, Kinlough-Rathbone RL, Mustard JF. Effect of repeated treatment of rabbit platelets with low concentrations of thrombin on their function, metabolism, and survival. Br J Haematol 1973; 25: 675-689
19 Booth WJ, Berndt MC, Castaldi PA. An altered platelet granule glycoprotein in patients with essential thrombocythemia. J Clin Invest 1984; 73: 291-297
20 Niewiarowski S, Budzynski AZ, Morinelli TA. Exposure of fibrinogen receptors on human platelets by proteolytic enzymes. J Biol Chem 1981; 256: 917-925
21 Zimmerman TS, Ratnoff OD, Littell AS. Detection of carriers of classic hemophilia using an immunologic assay for antihemophilia factor VIII). J Clin Invest 1971; 50: 244-254
22 Denson KW E, Biggs R. Human blood coagulation, haemostasis and thrombosis. Blackwell Sci Publ; Oxford: 1976: 325
23 Peake IR, Bloom AL, Giddings JC, Ludlam CA. An immunoradiometric assay for procoagulant factor VIII antigen: results in haemophilia, von Willebrand’s disease and fetal plasma and serum. Br J Haematol 1979; 42: 269-281
24 Frojmovie MM, Milton JG, Caen JP, Tobelem G. Platelets from “giant platelet syndrome (BSS)” are discocytes and normal sized. J Lab Clin Med 1978; 91: 109-116
5 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254
26 Scatchard G. The attraction of proteins for small molecules and ions. Ann NY Acad Sci 1949; 660-672
27 Neely SM, Gardner DV, Green D, Ts’ao C. Effect of hematin on endothelial cells and endothelial cell-platelet interactions. Am J Pathol 1984; 115: 390-396
28 Hoyer LW. The factor VIII complex: structure and function. Blood 1981; 58: 1-13