title: Disulphide bond assignment in human tissue inhibitor of metalloproteinases (TIMP).
abstract: Disulphide bonds in human recombinant tissue inhibitor of metalloproteinases (TIMP) were assigned by resolving proteolytic digests of TIMP on reverse-phase h.p.l.c. and sequencing those peaks judged to contain disulphide bonds by virtue of a change in retention time on reduction. This procedure allowed the direct assignment of Cys-145-Cys-166 and the isolation of two other peptides containing two disulphide bonds each. Further peptide cleavage in conjunction with fast-atom-bombardment m.s. analysis permitted the assignments Cys-1-Cys-70, Cys-3-Cys-99, Cys-13-Cys-124 and Cys-127-Cys-174 from these peptides. The sixth bond Cys-132-Cys-137 was assigned by inference, as the native protein has no detectable free thiol groups.
authors.affiliation: Biological Laboratory, University of Kent, Canterbury, U.K.
authors.forename: R A
authors.initials: RA
authors.lastname: Williamson
authors.forename: F A
authors.initials: FA
authors.lastname: Marston
authors.forename: S
authors.initials: S
authors.lastname: Angal
authors.forename: P
authors.initials: P
authors.lastname: Koklitis
authors.forename: M
authors.initials: M
authors.lastname: Panico
authors.forename: H R
authors.initials: HR
authors.lastname: Morris
authors.forename: A F
authors.initials: AF
authors.lastname: Carne
authors.forename: B J
authors.initials: BJ
authors.lastname: Smith
authors.forename: T J
authors.initials: TJ
authors.lastname: Harris
authors.forename: R B
authors.initials: RB
authors.lastname: Freedman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis; D004220:Disulfides / Q000032:analysis*; D006023:Glycoproteins; D006801:Humans; D006868:Hydrolysis; D008666:Metalloendopeptidases; D051379:Mice; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors; D008969:Molecular Sequence Data; D010434:Pepsin A; D011994:Recombinant Proteins / Q000235:genetics; D019714:Tissue Inhibitor of Metalloproteinases; D014357:Trypsin
chemical_list: D004220:Disulfides; D006023:Glycoproteins; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D004247:DNA; D014357:Trypsin; D010434:Pepsin A; D008666:Metalloendopeptidases; D003012:Microbial Collagenase