title: Substitution of gamma Arg-275 by Cys in an abnormal fibrinogen, "fibrinogen Osaka II". Evidence for a unique solitary cystine structure at the mutation site.
abstract: In an abnormal fibrinogen with impaired fibrin monomer polymerization designed as fibrinogen Osaka II, we have identified substitution of Arg by Cys at position 275 of the gamma chain. This Cys is linked to a free cysteine molecule by a disulfide link as evidenced by fast atom bombardment mass spectrometry. This finding was supported by identification of a single cysteine released from isolated abnormal fragment D1 upon reduction. This unique cystine structure at the mutation site has not been reported heretofore in any abnormal protein including fibrinogen. The substitution may well perturb the structure required for fibrin monomer polymerization, specifically that assigned to the carboxyl-terminal D domain of fibrinogen. Indeed, isolated fragment D1 with the Cys substitution failed to inhibit thrombin-mediated clotting of normal fibrinogen and normal fibrin monomer polymerization, while normal fragment D1 inhibited them markedly. Our data seem to provide supporting evidence that the putative polymerization site(s) assigned to the D domain of fibrinogen may be structure-dependent, including the carboxyl-terminal segment of the gamma chain as well as a contiguous region that contains the gamma 275 residue.
authors.affiliation: Division of Hemostasis and Thrombosis Research, Jichi Medical School, Tochigi-Ken, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Terukina
authors.forename: M
authors.initials: M
authors.lastname: Matsuda
authors.forename: H
authors.initials: H
authors.lastname: Hirata
authors.forename: Y
authors.initials: Y
authors.lastname: Takeda
authors.forename: T
authors.initials: T
authors.lastname: Miyata
authors.forename: T
authors.initials: T
authors.lastname: Takao
authors.forename: Y
authors.initials: Y
authors.lastname: Shimonishi
mesh_terms: D000293:Adolescent; D000368:Aged; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001120:Arginine; D001778:Blood Coagulation Disorders / Q000097:blood / Q000235:genetics*; D002118:Calcium / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine; D003553:Cystine; D004533:Egtazic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D015241:Fibrinogens, Abnormal; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D013918:Thrombin Time
chemical_list: D000596:Amino Acids; D015241:Fibrinogens, Abnormal; D010446:Peptide Fragments; C057129:fibrinogen Osaka II; D003553:Cystine; D004533:Egtazic Acid; D005337:Fibrin; D005340:Fibrinogen; D001120:Arginine; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase; D005341:Fibrinolysin; D003545:Cysteine; D002118:Calcium