title: Verification of protein disulfide bond arrangement by in-gel tryptic digestion under entirely neutral pH conditions.
abstract: To develop a concise proteomic procedure to verify the protein disulfide bond arrangement, non-reductive trypsin digestion of neuregulin 1-beta1 (176-246), a model disulfide-containing protein, was assessed by a proteolytic (18)O-labeling analysis. As a result, the commonly used in-gel tryptic digestion method has been improved for use entirely under neutral pH conditions. With this procedure, the disulfide arrangement of proteins could represent a clinical index candidate in pathological proteomic studies.
authors.affiliation: Laboratory of Proteomic Sciences, 21st Century COE Program, Kyoto Pharmaceutical University, Kyoto, Japan. saito@mb.kyoto-phu.ac.jp
authors.forename: Kazuki
authors.initials: K
authors.lastname: Saito
authors.forename: Itsuki
authors.initials: I
authors.lastname: Yasuo
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Uchimura
authors.forename: Shizuyo
authors.initials: S
authors.lastname: Koide-Yoshida
authors.forename: Takaaki
authors.initials: T
authors.lastname: Mizuguchi
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Kiso
mesh_terms: D000595:Amino Acid Sequence; D004220:Disulfides / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007553:Isotope Labeling / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020890:Neuregulin-1 / Q000737:chemistry / Q000378:metabolism; D010103:Oxygen Isotopes / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004220:Disulfides; D020890:Neuregulin-1; D010103:Oxygen Isotopes; D010446:Peptide Fragments; D014357:Trypsin