Immunologic studies of native and modified human factor VIII/von Willebrand factor.
Factor VIII/von Willebrand factor (FVIII/vWF) is a glycoprotein with a molecular weight greater than one-million daltons. Two activities are associated with this large molecule: FVIII procoagulant activity and vWF activity. Incubation of FVIII/vWF with proteolytic enzymes causes rapid inactivation of the FVIII procoagulant activity but has little effect on the vWF activity or antigenicity. In an attempt to gain insight into the structural features required for these two activities, antisera were raised in rabbits to normal, thrombin-inactivated, and plasmin-inactivated FVIII/vWF. All of these proteolytically modified forms of FVIII/vWF cross-reacted with each of the rabbit antisera; each blocked the ability of a human inhibitor to inactivate native active FVIII/vWF. Each of the antisera was a potent inhibitor of vWF activity and inactivated vWF activity at the same titer. The antisera were much less potent inhibitors of FVIII activity than of vWF activity. Antibodies to thrombin-inactivated FVIII/vWF or normal FVIII/vWF had about the same ability to inactivate FVIII procoagulant activity. Surprisingly, those to plasmin-inactivated FVIII/vWF still retained about 50% of this inhibitory capacity. A comparison of the three types of antigens by polyacrylamide gel electrophoresis in sodium dodecyl sulfate-6 M urea demonstrated that the structure of thrombin-inactivated FVIII/vWF was indistinguishable from that of normal FVIII/vWF, while plasmin-inactivated FVII/vWF was completely cleaved to lower molecular weight fragments. Some of the reported variations in the ability of rabbit antibodies to inhibit procoagulant activity may be due to partial degradation of the starting antigen. The retention by FVIII/vWF protein of its immunologic properties even after extensive proteolytic degradation suggests that under nondenaturing conditions, the conformation of the native and degraded molecules are very similar.
D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D001779:Blood Coagulation Factors / Q000276:immunology*; D003429:Cross Reactions; D004220:Disulfides; D000939:Epitopes; D005169:Factor VIII / Q000276:immunology*; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D008970:Molecular Weight; D013329:Structure-Activity Relationship; D013917:Thrombin / Q000378:metabolism; D014841:von Willebrand Factor / Q000276:immunology*
D001779:Blood Coagulation Factors; D004220:Disulfides; D000939:Epitopes; D014841:von Willebrand Factor; D005169:Factor VIII; D013917:Thrombin; D005341:Fibrinolysin