title: Structural model of factors V and Va based on scanning transmission electron microscope images and mass analysis.
abstract: Coagulation factor V (fV) is a single-chain glycoprotein (Mr 330,000; domain structure A1-A2-B-A3-C1-C2) that is activated to factor Va (fVa; Mr 174,000) by thrombin, which cleaves away the B domain leaving a heterodimeric structure composed of a heavy chain (A1-A2; Mr 94,000) and a light chain (A3-C1-C2; Mr 74,000). We analyzed the ultrastructure of scanning transmission electron microscope images of bovine and human fV, bovine fVa, and its constituent light chains and heavy chains. Factor V molecules had irregularly globular (10-12 nm) to oblong (8-14 nm) core structures which commonly displayed a peripheral satellite appendage of variable morphology attached to the core by a narrow stalk. Scanning transmission electron microscope mass analyses indicated that monomolecular bovine fV molecules had a mass of 322 +/- 45 kDa and human fV, 315 +/- 31 kDa. Factor Va molecules were irregular, globular (8-12 nm) structures that resembled the fV core structure, lacked the satellite appendage representing B domainal structures, and had a mass of 180 +/- 22 kDa. Our findings permit us to propose a structural model of fV suggesting the relative orientation of its closely associated light chain and heavy chain core components and indicating that these constituents remain associated in the transition from fV to fVa.
authors.affiliation: University of Wisconsin Medical School-Milwaukee Clinical Campus, Milwaukee 53233.
authors.forename: M W
authors.initials: MW
authors.lastname: Mosesson
authors.forename: W R
authors.initials: WR
authors.lastname: Church
authors.forename: J P
authors.initials: JP
authors.lastname: DiOrio
authors.forename: S
authors.initials: S
authors.lastname: Krishnaswamy
authors.forename: K G
authors.initials: KG
authors.lastname: Mann
authors.forename: J F
authors.initials: JF
authors.lastname: Hainfeld
authors.forename: J S
authors.initials: JS
authors.lastname: Wall
mesh_terms: D000818:Animals; D002417:Cattle; D005165:Factor V / Q000648:ultrastructure*; D015943:Factor Va / Q000648:ultrastructure*; D006801:Humans; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D008958:Models, Molecular; D008970:Molecular Weight; D011487:Protein Conformation
chemical_list: D046911:Macromolecular Substances; D015943:Factor Va; D005165:Factor V