Using a Poisson approximation to predict the isotopic distribution of sulphur-containing peptides in a peptide-centric proteomic approach.

Breen et al. (Electrophoresis 2000; 21: 2243) proposed a method for finding monoisotopic peptide peaks in mass spectra based on an approximation of the distribution of different isotopic variants of a peptide by a Poisson distribution. They developed the method using all protein sequences from the SWISS-PROT database. We investigate the suitability of this method to predict the isotopic distribution in an environment which enriches for peptides carrying sulphur. More specifically, we focus on mass spectra obtained by a COmbined FRActional DIagonal Chromatography (COFRADIC) approach, developed by Gevaert et al. (Nature Biotechnology 2003; 21: 566), targeting a specific subset of peptides, in this case the N-terminal peptides. One can therefore ask whether the original results of Breen et al. apply to spectra generated by the particular COFRADIC method. We investigate whether the proposed approximation holds for N-terminal peptides. We also evaluate whether ignoring sulphur atoms while developing the approximation, as proposed by Breen et al., does not increase the risk of missing monoisotopic peaks corresponding to sulphur-containing peptides. Finally, we check the sensitivity of the quality of the approximation to optimization criteria used in the development process. The results are not simply restricted to a COFRADIC setting but are also applicable more generally, for any method which enriches for sulphur-containing peptides. Copyright (c) 2007 John Wiley & Sons, Ltd.