title: Antithrombin-III-Stockholm: a codon 392 (Gly----Asp) mutation with normal heparin binding and impaired serine protease reactivity.
abstract: Antithrombin-III-Stockholm is a new structural variant of antithrombin-III (AT-III) with normal heparin affinity but defective serine protease inhibitory activity. The proposita, a white female born in 1966, was diagnosed to have developed a pulmonary embolus while on oral contraceptives at age 19. The proposita, as well as her father, were diagnosed to have a type 2 AT-III deficiency as they had normal levels of immunoreactive AT-III associated with decreased (approximately 60%) functional AT-III when measured with either alpha-thrombin or factor Xa as the substrate, either in the presence or absence of heparin. There was no evidence of abnormal electrophoretic mobility of AT-III from the proposita either in the presence or absence of heparin. Genomic DNA was prepared and all seven AT-III exons were polymerase chain reaction (PCR)-amplified and sequenced in both directions using nested primers. Only exon 7 provided evidence for the presence of a mutation, with the second base of codon 392 having a G----A substitution. Such a mutation would cause the substitution of aspartic acid at the site of the normally appearing glycine in the translated product. Furthermore, this mutation caused the destruction of an Hae III restriction site at this point in the AT-III gene. The absence of this Hae III site was confirmed using restriction fragment length polymorphism analysis of PCR-amplified material from the proposita. Experiments with AT-III from the proposita together with experiments with cell-free translated AT-III-Stockholm provided evidence that the mutant AT-III protein does not efficiently form a stable covalent inhibitory complex with alpha-thrombin, although it exhibits normal heparin affinity. The minimal thrombin-complexing ability of the mutant AT-III protein that was observed was accelerated by heparin, but to subnormal levels.
authors.affiliation: Canadian Red Cross Blood Society Transfusion Service, Hamilton, Ontario.
authors.forename: M A
authors.initials: MA
authors.lastname: Blajchman
authors.forename: F
authors.initials: F
authors.lastname: Fernandez-Rachubinski
authors.forename: W P
authors.initials: WP
authors.lastname: Sheffield
authors.forename: R C
authors.initials: RC
authors.lastname: Austin
authors.forename: S
authors.initials: S
authors.lastname: Schulman
mesh_terms: D000328:Adult; D000990:Antithrombin III / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003062:Codon; D005091:Exons; D005260:Female; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000032:analysis*; D013917:Thrombin / Q000378:metabolism
chemical_list: D003062:Codon; C073583:antithrombin III Stockholm; D000990:Antithrombin III; D006493:Heparin; D012697:Serine Endopeptidases; D013917:Thrombin