title: Characterization of human factor VIII and interaction with von Willebrand factor. An electron microscopic study.
abstract: Blood coagulation factor VIII is a large glycoprotein that circulates in plasma at relative low concentration (0.1 microgram/ml). It consists of a heterogeneous mixture of a series heavy-chain peptides (90-200 kDa), each associated with a light chain of 80 kDa. To gain insight into the physical properties of the protein, we have characterized purified human factor VIII by electron microscopy and rotary shadowing. Electron microscopy of rotary shadowed factor VIII molecules showed predominantly a single globular domain structure, with a somewhat asymmetric shape, while two-domain structures were also encountered. The overall dimensions of the globular domains ranged from 4 x 6 nm to 8 x 12 nm. EDTA treatment of factor VIII reduced the overall dimensions (2.5 x 5 nm to 6 x 10 nm) while treatment with thrombin reduced the dimensions to a small extent. In complexes with von Willebrand factor, factor VIII appeared localized at the globular domains of von Willebrand factor multimers. In addition, incubation of factor VIII with Staphylococcus aureus V8 protease fragments SpII and SpIII revealed only binding to the globular domains of SpIII. In this study, the first morphological characterization of human factor VIII is presented, together with its direct localization on von Willebrand factor multimers.
authors: Department of Cell Biology, Medical Faculty, University of Utrecht, The Netherlands.
authors: H F
authors: HF
authors: Heijnen
authors: J A
authors: JA
authors: Koedam
authors: H
authors: H
authors: Sandberg
authors: N H
authors: NH
authors: Beeser-Visser
authors: J W
authors: JW
authors: Slot
authors: J J
authors: JJ
authors: Sixma
mesh_terms: D015153:Blotting, Western; D002850:Chromatography, Gel; D004492:Edetic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D005169:Factor VIII / Q000378:metabolism / Q000648:ultrastructure*; D006801:Humans; D008854:Microscopy, Electron; D013917:Thrombin; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D014841:von Willebrand Factor; D005169:Factor VIII; D004492:Edetic Acid; D013917:Thrombin