title: Protein S Tokushima: abnormal molecule with a substitution of Glu for Lys-155 in the second epidermal growth factor-like domain of protein S.
abstract: A 29-year-old female patient with heterozygous congenital protein S deficiency suffering from thrombotic disease had normal levels of both total and free protein S antigen (70% and 65%, respectively), but low cofactor activity (31%) for activated protein C, indicating that she had a variant of protein S, protein S Tokushima. Western blotting using the polyclonal anti-protein S antibody showed that approximately half of the patient's protein S appeared to be the variant with a higher molecular weight than normal protein S. The partially purified variant protein S bound neither to the monoclonal antibody recognizing calcium-dependent conformation of protein S nor to the antibody recognizing the thrombin-sensitive domain of protein S. Among the exons from II to XV of the patient's protein S gene encoding from the NH2-terminal end to the COOH-terminal end of protein S, only one missense mutation (A to G) was found in exon VI of the protein S alpha-gene, which results in amino acid substitution of Glu(GAG) for Lys-155(AAG) in the second epidermal growth factor-like domain of protein S. The recombinant protein S Tokushima expressed in BHK cells had a slightly higher molecular weight than the recombinant normal one, did not bind to the antibody specific for the thrombin-sensitive domain, and did not show the cofactor activity. These findings suggest that the protein S Tokushima molecule is structurally and functionally a variant of protein S, and that this variant protein S is the cause of severe thrombosis in this patient.
authors.affiliation: Department of Molecular Biology on Genetic Disease, Mie University School of Medicine, Tsu-city, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Hayashi
authors.forename: J
authors.initials: J
authors.lastname: Nishioka
authors.forename: T
authors.initials: T
authors.lastname: Shigekiyo
authors.forename: S
authors.initials: S
authors.lastname: Saito
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D004815:Epidermal Growth Factor / Q000737:chemistry; D005091:Exons; D005260:Female; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D017293:Protein S / Q000737:chemistry / Q000235:genetics*; D018455:Protein S Deficiency; D011994:Recombinant Proteins / Q000737:chemistry; D013927:Thrombosis / Q000235:genetics
chemical_list: D017293:Protein S; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor