Journal
PROTEOMICS
Volume 10, Issue 8, Pages 1545-1556Publisher
WILEY
DOI: 10.1002/pmic.200900255
Keywords
Bioinformatics; Database search; iTRAQ; Quantitation
Funding
- Ramon y Cajal [RYC-2006-001446, RYC-2007-00228]
- Fundacao para a Ciencia e a Tecnologia, Portugal [C2007-IPATIMUP/AA2]
- Programa Operacional Ciencia e Inovacao 2010 [POCI 2010]
- VI Programa Quadro [2002-2006]
- Spanish Ministry of Health [06/0621]
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A frequent goal of MS-based proteomics experiments nowadays is to quantify changes in the abundance of proteins across several biological samples. The iTRAQ labeling method is a powerful technique; when combined with LC coupled to MS/MS it allows relative quantitation of up to eight different samples simultaneously. Despite the usefulness of iTRAQ current software solutions have limited functionality and require the combined use of several software programs for analysis of the data from different MS vendors. We developed an integrated tool, now available in the virtual expert mass spectrometrist (VEMS) program, for database-dependent search of MS/MS spectra, quantitation and database storage for iTRAQ-labeled samples. VEMS also provides useful alternative report types for large-scale quantitative experiments. The implemented statistical algorithms build on quantitative algorithms previously used in proposed iTRAQ tools as described in detail herein. We propose a new algorithm, which provides more accurate peptide ratios for data that show an intensity-dependent saturation. The accuracy of the proposed iTRAQ algorithm and the performance of VEMS are demonstrated by comparing results from VEMS, MASCOT and PEAKS Q obtained by analyzing data from a reference mixture of six proteins. Users can download VEMS and test data from http://www.portugene.com/software.html.
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