An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database-英文文献.pdf

An Approach to Correlate Tandem Mass Spectral Data of Peptides with Amino Acid Sequences in a Protein Database Jimmy K. Eng, Ashley L. McCormack, and John R. Yates, III Departmentof Molecular Biotechnology,University of Washington,Seattle, Washington,USA A method to correlate the uninterpreted tandem mass spectra of peptides produced under low energy (lo-50 eV) collision conditions with amino acid sequences in the Genpept database has been developed. In this method the protein database is searched to identify linear amino acid sequences within a mass tolerance of * 1 u of the precursor ion molecular weight. A cross-correlation function is then used to provide a measurement of similarity between the mass-to-charge ratios for the fragment ions predicted from amino acid se- quences obtained from the database and the fragment ions observed in the tandem mass spectrum. In general, a difference greater than 0.1 between the normalized cross-correlation functions of the first- and second-ranked search results indicates a successfol match between sequence and spectrum. Searches of species-specific protein databases with tandem mass spectra acquired from peptides obtained from the enzymatically digested total proteins of E. coli and S. cerevisiae cells allowed matchmg of the spectra to amino acid sequences within proteins of these organisms. The approach described in this manuscript provides a conve- nient method to interpret tandem mass spectra w