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Reverse and Random Decoy Methods for False Discovery Rate Estimation in High Mass Accuracy Peptide Spectral Library Searches

Spectral library searching (SLS) is an attractive alternative to sequence database searching (SDS) for peptide identification due to its speed, sensitivity, and ability to include any selected mass spectra. While decoy methods for SLS have been developed for low mass accuracy peptide spectral libraries, it is not clear that they are optimal or directly applicable to high mass accuracy spectra. Therefore, we report the development and validation of methods for high mass accuracy decoy libraries. Two types of decoy libraries were found to be suitable for this purpose. The first, referred to as Reverse, constructs spectra by reversing a library’s peptide sequences except for the C-terminal residue. The second, termed Random, randomly replaces all non-C-terminal residues and either retains the original C-terminal residue or replaces it based on the amino-acid frequency of the library’s C-terminus. In both cases the m/z values of fragment ions are shifted accordingly. Determination of FDR is performed in a man...

Autoren:   Zheng Zhang; Meghan Burke; Yuri A. Mirokhin; Dmitrii V. Tchekhovskoi; Sanford P. Markey; Wen Yu; Raghothama Chaerkady; Sonja Hess; Stephen E. Stein
Journal:   Journal of Proteome Research
Jahrgang:   2018
DOI:   10.1021/acs.jproteome.7b00614
Erscheinungsdatum:   11.01.2018
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