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Proteomics. 2010 Jun;10(12):2369-76. doi: 10.1002/pmic.200900619.

Estimating false discovery rates for peptide and protein identification using randomized databases.

Author information

1
Bioinformatics & High-throughput Analysis Laboratory, Seattle Children's Research Institute, Seattle, WA 98101, USA.

Abstract

MS-based proteomics characterizes protein contents of biological samples. The most common approach is to first match observed MS/MS peptide spectra against theoretical spectra from a protein sequence database and then to score these matches. The false discovery rate (FDR) can be estimated as a function of the score by searching together the protein sequence database and its randomized version and comparing the score distributions of the randomized versus nonrandomized matches. This work introduces a straightforward isotonic regression-based method to estimate the cumulative FDRs and local FDRs (LFDRs) of peptide identification. Our isotonic method not only performed as well as other methods used for comparison, but also has the advantages of being: (i) monotonic in the score, (ii) computationally simple, and (iii) not dependent on assumptions about score distributions. We demonstrate the flexibility of our approach by using it to estimate FDRs and LFDRs for protein identification using summaries of the peptide spectra scores. We reconfirmed that several of these methods were superior to a two-peptide rule. Finally, by estimating both the FDRs and LFDRs, we showed for both peptide and protein identification, moderate FDR values (5%) corresponded to large LFDR values (53 and 60%).

PMID:
20391536
DOI:
10.1002/pmic.200900619
[Indexed for MEDLINE]

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