title: Characterization of a factor VII molecule carrying a mutation in the second epidermal growth factor-like domain.
abstract: A missense mutation at codon 100 in the second epidermal growth factor-like domain, resulting in Gln100-->Arg, was detected in 19 out of 21 available severely factor VII (FVII) deficient patients in Norway. Seventeen patients were homozygous, and the two remaining were compound heterozygotes. In the homozygous patients, FVII antigen was measured to 10-28%, and activity to 0.6-6.5% of that in normal pooled plasma. Recombinant FVII containing the mutation was expressed transiently in CHO cells to a mean antigen level of 57% of the wild type FVII protein, and with a specific activity of 6% of wild type. The mutant protein had a 14-fold reduction in affinity for tissue factor (TF), whereas binding of FX seemed unaffected. In line with the experimental data, molecular modelling of the mutation based on the coordinates of the tissue factor/FVIIa complex showed that substituting arginine for glutamine disrupts the interface between the catalytic and second epidermal growth factor-like domains.
authors.affiliation: Biotechnology Centre of Oslo, University of Oslo, Norway.
authors.forename: A
authors.initials: A
authors.lastname: Kavlie
authors.forename: L
authors.initials: L
authors.lastname: Orning
authors.forename: A
authors.initials: A
authors.lastname: Grindflek
authors.forename: H
authors.initials: H
authors.lastname: Stormorken
authors.forename: H
authors.initials: H
authors.lastname: Prydz
mesh_terms: D000293:Adolescent; D000328:Adult; D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001780:Blood Coagulation Tests; D016466:CHO Cells; D002648:Child; D002675:Child, Preschool; D003062:Codon / Q000235:genetics*; D006224:Cricetinae; D004252:DNA Mutational Analysis; D004789:Enzyme Activation; D005167:Factor VII / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005168:Factor VII Deficiency / Q000208:ethnology / Q000235:genetics*; D005170:Factor X / Q000378:metabolism; D005260:Female; D005838:Genotype; D005973:Glutamine / Q000737:chemistry; D006801:Humans; D008297:Male; D008875:Middle Aged; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009664:Norway / Q000453:epidemiology; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis; D013925:Thromboplastin / Q000378:metabolism; D014162:Transfection
chemical_list: D003062:Codon; D011994:Recombinant Proteins; D005973:Glutamine; D005167:Factor VII; D005170:Factor X; D013925:Thromboplastin