title: Electron crystallography of human blood coagulation factor VIII bound to phospholipid monolayers.
abstract: Coagulation factor VIII binds to negatively charged platelets prior to assembly with the serine protease, factor IXa, to form the factor X-activating enzyme (FX-ase) complex. The macromolecular organization of membrane-bound factor VIII has been studied by electron crystallography for the first time. For this purpose two-dimensional crystals of human factor VIII were grown onto phosphatidylserine-containing phospholipid monolayers, under near to physiological conditions (pH and salt concentration). Electron crystallographic analysis revealed that the factor VIII molecules were organized as monomers onto the lipid layer, with unit cell dimensions: a = 81.5A, b = 67.2 A, gamma = 66.5 degrees, P1 symmetry. Based on a homology-derived molecular model of the factor VIII (FVIII) A domains, the FVIII projection structure solved at 15-A resolution presents the A1, A2, and A3 domain heterotrimer tilted approximately 65 degrees relative to the membrane plane. The A1 domain is projecting on top of the A3, C1, and C2 domains and with the A2 domain protruding partially between A1 and A3. This organization of factor VIII allows the factor IXa protease and epidermal growth factor-like domain binding sites (localized in the A2 and A3 domains, respectively) to be situated at the appropriate position for the binding of factor IXa. The conformation of the lipid-bound FVIII is therefore very close to that for the activated factor VIIIa predicted in the FX-ase complex.
authors.affiliation: School of Biochemistry and Molecular Biology, The University of Leeds, Leeds LS2 9JT, United Kingdom. svetla@bmb.leeds.ac.uk
authors.forename: S S
authors.initials: SS
authors.lastname: Stoylova
authors.forename: P J
authors.initials: PJ
authors.lastname: Lenting
authors.forename: G
authors.initials: G
authors.lastname: Kemball-Cook
authors.forename: A
authors.initials: A
authors.lastname: Holzenburg
mesh_terms: D003461:Crystallography; D005169:Factor VIII / Q000737:chemistry*; D006801:Humans; D010743:Phospholipids; D011487:Protein Conformation
chemical_list: D010743:Phospholipids; D005169:Factor VIII