title: Characterization of dysfunctional factor VIII molecules.
abstract: Immunopurification and characterization of dysfunctional factor VIII-like molecules in CRM-positive and CRM-reduced hemophilia A permit correlation of structural changes with molecular defects. The technique described here is sufficiently sensitive to characterize the molecular mass and enzymatic fragments of the factor VIII chains in patients with as little VIII: Ag as 0.05 units/ml. Specific abnormalities have been identified in 5 of the first 24 samples tested. In each case, the mutation responsible for factor VIII dysfunction has been determined by sequencing a part of the abnormal gene. Mutations have been identified that abolish critical thrombin cleavage sites or which generate new N-glycosylation sites. The technique provides a useful approach to the study factor VIII structure-function relationships, and it has the potential to clarify further the molecular basis of factor VIII procoagulant activity.
authors.affiliation: Holland Laboratory, American Red Cross, Rockville, Maryland 20855.
authors.forename: L W
authors.initials: LW
authors.lastname: Hoyer
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004589:Electrophoresis, Disc / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005169:Factor VIII / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D006031:Glycosylation; D006467:Hemophilia A / Q000097:blood / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007163:Immunosorbent Techniques; D007202:Indicators and Reagents; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes / Q000302:isolation & purification; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D013917:Thrombin / Q000378:metabolism
chemical_list: D007202:Indicators and Reagents; D009692:Nucleic Acid Heteroduplexes; D010446:Peptide Fragments; D005169:Factor VIII; D004247:DNA; D013917:Thrombin