Fast liquid chromatography coupled to electrospray tandem mass spectrometry peptide sequencing for cross-species protein identification.
Using a parallel microcolumn switching liquid chromatography set-up coupled to a quadrupole time-of-flight mass spectrometer, a rapid liquid chromatography/mass spectrometric (LC/MS) protein identification method is presented. Without prior sample clean-up up to 300 protein digest samples a day can be processed. Using data-directed acquisition, up to 10 fragmentation analyses for each protein sample can be acquired in the same chromatographic run that can be used for database searching. Using internal peptide sequence information, protein databases and the various nucleic acid databases can both be queried for cross-species identification of the protein sample. The method was evaluated and put into force to generate data for a tobacco cell culture protein database.
D002460:Cell Line; D002853:Chromatography, Liquid; D030562:Databases, Protein; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D020539:Sequence Analysis, Protein / Q000379:methods*; D013045:Species Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors; D014026:Nicotiana / Q000737:chemistry* / Q000166:cytology
D011506:Proteins