title: Factor V turnover in a primate model.
abstract: The isolation and characterization of baboon plasma factor V (FV) were performed for the development of an in vivo model for studying factor V/Va physiology in nonhuman primates. Baboon FV was purified by immunoaffinity chromatography with an antihuman FV monoclonal antibody and exhibits a specific activity of 1,940 U/mg. Baboon FV activation by thrombin proceeds through two proteolytic pathways similar to those observed with human and bovine FV. Limited amino acid sequencing of FV and its thrombin activation fragments shows 95% identity with human and 79% identity with bovine FV. 125I-Factor V and a mixture of thrombin cleaved 125I-FV activation products were infused into normal male baboons and evaluated by blood sample radioactivity measurements and by autoradiography of plasma samples following resolution by gel electrophoresis. Factor V disappeared with a half-life (t1/2) of 12.98 +/- 1.85 hours and was cleared without obvious degradation of the molecule during circulation. The radioactivity associated with the thrombin activated FV mixture, which consisted of the Mr = 220,000 activation intermediate, the Mr = 150,000 activation peptide, the heavy chain (HC) and the light chain (LC) of FVa, was cleared in a nonlinear manner. The HC and LC were removed with t1/2 < 20 minutes. The apparent molecular weight (Mr) = 220,000 and Mr = 150,000 fragments were cleared with t1/2 > 6 hours and t1/2 > 30 hours, respectively.
authors.affiliation: Department of Biochemistry, University of Vermont, College of Medicine, Burlington 05405-0068, USA.
authors.forename: M D
authors.initials: MD
authors.lastname: Rand
authors.forename: S R
authors.initials: SR
authors.lastname: Hanson
authors.forename: K G
authors.initials: KG
authors.lastname: Mann
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001345:Autoradiography; D002846:Chromatography, Affinity / Q000706:statistics & numerical data; D004591:Electrophoresis, Polyacrylamide Gel; D005165:Factor V / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D015943:Factor Va / Q000378:metabolism; D006801:Humans; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D007700:Kinetics; D008297:Male; D008657:Metabolic Clearance Rate; D008954:Models, Biological; D010215:Papio / Q000097:blood; D011486:Protein C / Q000378:metabolism; D012680:Sensitivity and Specificity; D017421:Sequence Analysis
chemical_list: D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes; D011486:Protein C; D015943:Factor Va; D005165:Factor V