title: Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels.
abstract: Proteins from silver-stained gels can be digested enzymatically and the resulting peptide analyzed and sequenced by mass spectrometry. Standard proteins yield the same peptide maps when extracted from Coomassie- and silver-stained gels, as judged by electrospray and MALDI mass spectrometry. The low nanogram range can be reached by the protocols described here, and the method is robust. A silver-stained one-dimensional gel of a fraction from yeast proteins was analyzed by nano-electrospray tandem mass spectrometry. In the sequencing, more than 1000 amino acids were covered, resulting in no evidence of chemical modifications due to the silver staining procedure. Silver staining allows a substantial shortening of sample preparation time and may, therefore, be preferable over Coomassie staining. This work removes a major obstacle to the low-level sequence analysis of proteins separated on polyacrylamide gels.
authors.affiliation: European Molecular Biology laboratory (EMBL), Heidelberg, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Shevchenko
authors.forename: M
authors.initials: M
authors.lastname: Wilm
authors.forename: O
authors.initials: O
authors.lastname: Vorm
authors.forename: M
authors.initials: M
authors.lastname: Mann
mesh_terms: D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis*; D017421:Sequence Analysis / Q000379:methods*; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D007202:Indicators and Reagents; D011506:Proteins