Protein C Is Responsible for the Rapid Inactivation of Factor Va following Blood Clotting In Vitro

Denise E Jackson; Christina A Mitchell; Hatem H Salem

doi:10.1055/s-0038-1648813

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1994; 72(01): 070-073
DOI: 10.1055/s-0038-1648813

Original Article

Schattauer GmbH Stuttgart

Protein C Is Responsible for the Rapid Inactivation of Factor Va following Blood Clotting In Vitro

Denise E Jackson

The Department of Medicine, Monash University, Box Hill Hospital, Box Hill, Victoria, Australia

,

Christina A Mitchell

The Department of Medicine, Monash University, Box Hill Hospital, Box Hill, Victoria, Australia

,

Hatem H Salem

The Department of Medicine, Monash University, Box Hill Hospital, Box Hill, Victoria, Australia

› Author Affiliations

Abstract

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Summary

When whole blood is allowed to clot in vitro, factor V is rapidly activated to factor Va which is subsequently inactivated. We developed two monoclonal anti-protein C antibodies, one of which inhibits protein C activation and the other inhibits protein C activity. The addition of either antibody to blood before clotting in vitro significantly inhibited the inactivation of factor Va, confirming the essential role of protein C in mediating the rapid inactivation of factor Va.

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References

References
1 Salem HH, Broze GJ, Miletich JP, Majerus PW. Human coagulation factor Va is a cofactor for the activation of protein C. Proc Natl Acad Sci 1983; 80: 1584-1588
2 Suzuki K, Dahlback B, Stenflo J. Thrombin-catalyzed activation of human coagulation factor V. J Biol Chem 1982; 267: 6556-6564
3 Seegers WH. Comments on Ac-Globulin (Factor V): A source of personal rewards. Thromb Res 1974; 5: 703-707
4 Mertens K, Bertina RM. Pathways in the activation of human coagulation factor X. Biochem J 1980; 185: 647-658
15 Laemmli UK. Cleavage of structural protein during the assembly on the head of the bacteriophage T4. Nature 1970; 227: 680-685
6 Thompson EA, Salem HH. Modification of human thrombin: effect on thrombomodulin binding. Thromb Haemost 1988; 59: 415-420
7 Owen WG. Evidence for the formation of an ester between thrombin and heparin cofactor. Biochem Biophys Acta 1975; 405: 380-387
8 Dahlback B. Purification of human vitamin K dependent protein S and its limited proteolysis by thrombin. Biochem J 1983; 209: 837-846
9 Jakobsen E, Koerulf P. A modified beta alanine precipitation procedure to prepare fibrinogen free of anti-thrombin III and plasminogen. Thromb Res 1973; 3: 145-149
10 Salem HH, Maruyama I, Ishii H, Majerus PW. Isolation and characterization of thrombomodulin from human placenta. J Biol Chem 1984; 259: 12246-12251
11 McEver RP, Baenziger NL, Majerus PW. Isolation and quantitation of the platelet glycoprotein deficient in thrombasthenia using a monoclonal hybridoma antibody. J Clin Invest 1980; 66: 1311-1318
12 Harlow E, Lane D. (eds). Antibodies - A laboratory manual. New York, NY: Cold Spring Harbor Laboratory; 1988: 288-304
13 Burnette WN. “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulphate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 1981; 112: 195-204
14 Beesley JE. (ed). Colloidal gold: A new perspective for cytochemical marking. Microscopy Handbooks, 17 Oxford University Press; England: 1989: 1-58
15 Miletich JP, Majerus DW, Majerus PW. Patients with congenital factor V deficiency have decreased factor Xa binding sites on their platelets. J Clin Invest 1978; 62: 826-831
16 Oates AM, Salem HH. The regulation of human factor V by a neutrophil protease. Blood 1987; 70: 846-851