Premium
Archived polyacrylamide gels as a resource for proteome characterization by mass spectrometry
Author(s) -
Shevchenko Anna,
Loboda Alexander,
Ens Werner,
Schraven Burkhart,
Standing Kenneth G.,
Shevchenko Andrej
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683()22:6<1194::aid-elps1194>3.0.co;2-a
Subject(s) - proteome , mass spectrometry , chemistry , protein mass spectrometry , chromatography , tandem mass spectrometry , bottom up proteomics , matrix assisted laser desorption/ionization , proteomics , peptide , polyacrylamide gel electrophoresis , gel electrophoresis , two dimensional gel electrophoresis , tandem mass tag , desorption , biochemistry , quantitative proteomics , enzyme , organic chemistry , adsorption , gene
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.