title: Archived polyacrylamide gels as a resource for proteome characterization by mass spectrometry.
abstract: Mass spectrometry was applied to identify protein spots excised from an archived two-dimensional polyacrylamide gel that had been dried and stored for eight years at room temperature. All proteins were successfully identified. Detailed characterization of protein digests by matrix-assisted laser desorption/ionization (MALDI) peptide mapping, nanoelectrospray tandem mass spectrometry and MALDI-quadrupole time-of-flight mass spectrometry revealed no evidence of protein degradation or modifications that could hamper identification of proteins in a sequence database. The experiment with a model protein demonstrated that the pattern of tryptic peptides and the yield of individual peptides were not noticeably changed in the in-gel digest of the archived protein spot compared to the digest of the spot excised from a fresh gel. Thus, the characterization of "archived proteomes" has the potential to advance proteomic research without repeating "wet" biochemistry experiments, that had been perfected in the laboratory years ago.
authors.affiliation: Peptide and Protein Group, European Molecular Biology Laboratory, Heidelberg, Germany. schevchenko@EMBL-Heidelberg.de
authors.forename: A
authors.initials: A
authors.lastname: Shevchenko
authors.forename: A
authors.initials: A
authors.lastname: Loboda
authors.forename: W
authors.initials: W
authors.lastname: Ens
authors.forename: B
authors.initials: B
authors.lastname: Schraven
authors.forename: K G
authors.initials: KG
authors.lastname: Standing
authors.forename: A
authors.initials: A
authors.lastname: Shevchenko
mesh_terms: D000180:Acrylic Resins; D000595:Amino Acid Sequence; D006801:Humans; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014357:Trypsin / Q000378:metabolism
chemical_list: D000180:Acrylic Resins; D020543:Proteome; C016680:polyacrylamide gels; D014357:Trypsin