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      PRIDE Inspector Toolsuite: Moving Toward a Universal Visualization Tool for Proteomics Data Standard Formats and Quality Assessment of ProteomeXchange Datasets*

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          Abstract

          The original PRIDE Inspector tool was developed as an open source standalone tool to enable the visualization and validation of mass-spectrometry (MS)-based proteomics data before data submission or already publicly available in the Proteomics Identifications (PRIDE) database. The initial implementation of the tool focused on visualizing PRIDE data by supporting the PRIDE XML format and a direct access to private (password protected) and public experiments in PRIDE.

          The ProteomeXchange (PX) Consortium has been set up to enable a better integration of existing public proteomics repositories, maximizing its benefit to the scientific community through the implementation of standard submission and dissemination pipelines. Within the Consortium, PRIDE is focused on supporting submissions of tandem MS data. The increasing use and popularity of the new Proteomics Standards Initiative (PSI) data standards such as mzIdentML and mzTab, and the diversity of workflows supported by the PX resources, prompted us to design and implement a new suite of algorithms and libraries that would build upon the success of the original PRIDE Inspector and would enable users to visualize and validate PX “complete” submissions. The PRIDE Inspector Toolsuite supports the handling and visualization of different experimental output files, ranging from spectra (mzML, mzXML, and the most popular peak lists formats) and peptide and protein identification results (mzIdentML, PRIDE XML, mzTab) to quantification data (mzTab, PRIDE XML), using a modular and extensible set of open-source, cross-platform libraries. We believe that the PRIDE Inspector Toolsuite represents a milestone in the visualization and quality assessment of proteomics data. It is freely available at http://github.com/PRIDE-Toolsuite/.

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          Most cited references33

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          The Proteomics Identifications (PRIDE) database and associated tools: status in 2013

          The PRoteomics IDEntifications (PRIDE, http://www.ebi.ac.uk/pride) database at the European Bioinformatics Institute is one of the most prominent data repositories of mass spectrometry (MS)-based proteomics data. Here, we summarize recent developments in the PRIDE database and related tools. First, we provide up-to-date statistics in data content, splitting the figures by groups of organisms and species, including peptide and protein identifications, and post-translational modifications. We then describe the tools that are part of the PRIDE submission pipeline, especially the recently developed PRIDE Converter 2 (new submission tool) and PRIDE Inspector (visualization and analysis tool). We also give an update about the integration of PRIDE with other MS proteomics resources in the context of the ProteomeXchange consortium. Finally, we briefly review the quality control efforts that are ongoing at present and outline our future plans.
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            TANDEM: matching proteins with tandem mass spectra.

            Tandem mass spectra obtained from fragmenting peptide ions contain some peptide sequence specific information, but often there is not enough information to sequence the original peptide completely. Several proprietary software applications have been developed to attempt to match the spectra with a list of protein sequences that may contain the sequence of the peptide. The application TANDEM was written to provide the proteomics research community with a set of components that can be used to test new methods and algorithms for performing this type of sequence-to-data matching. The source code and binaries for this software are available at http://www.proteome.ca/opensource.html, for Windows, Linux and Macintosh OSX. The source code is made available under the Artistic License, from the authors.
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              A probability-based approach for high-throughput protein phosphorylation analysis and site localization.

              Data analysis and interpretation remain major logistical challenges when attempting to identify large numbers of protein phosphorylation sites by nanoscale reverse-phase liquid chromatography/tandem mass spectrometry (LC-MS/MS) (Supplementary Figure 1 online). In this report we address challenges that are often only addressable by laborious manual validation, including data set error, data set sensitivity and phosphorylation site localization. We provide a large-scale phosphorylation data set with a measured error rate as determined by the target-decoy approach, we demonstrate an approach to maximize data set sensitivity by efficiently distracting incorrect peptide spectral matches (PSMs), and we present a probability-based score, the Ascore, that measures the probability of correct phosphorylation site localization based on the presence and intensity of site-determining ions in MS/MS spectra. We applied our methods in a fully automated fashion to nocodazole-arrested HeLa cell lysate where we identified 1,761 nonredundant phosphorylation sites from 491 proteins with a peptide false-positive rate of 1.3%.
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                Author and article information

                Journal
                Mol Cell Proteomics
                Mol. Cell Proteomics
                mcprot
                mcprot
                MCP
                Molecular & Cellular Proteomics : MCP
                The American Society for Biochemistry and Molecular Biology
                1535-9476
                1535-9484
                January 2016
                6 November 2015
                6 November 2015
                : 15
                : 1
                : 305-317
                Affiliations
                [1]From the ‡European Molecular Biology Laboratory, European Bioinformatics Institute (EMBL-EBI), Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SD, UK;
                [2]§Ruhr-Universität Bochum, Medizinisches Proteom-Zenter, Medical Bioinformatics, ZKF, E.142, Universitätsstr. 150, D-44801 Bochum, Germany;
                [3]¶Division of Immunology, Allergy and Infectious Diseases, Department of Dermatology, Medical University of Vienna, Austria;
                [4]‖Department of Proteomics, Center for Genetic Engineering and Biotechnology, Ciudad de la Habana, Cuba
                Author notes
                ** To whom correspondence should be addressed: European Molecular Biology Laboratory, European Bioinformatics Institute (EMBL-EBI), Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SD, UK. Tel.: +44 1223 492 610; Fax: +44 1223 494 484; E-mail: juan@ 123456ebi.ac.uk .
                Article
                O115.050229
                10.1074/mcp.O115.050229
                4762524
                26545397
                e00919db-2fc6-48b2-9a83-7a4e627d3c3e
                © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

                Author's Choice—Final version free via Creative Commons CC-BY license.

                History
                : 20 April 2015
                : 4 November 2015
                Funding
                Funded by: Biotechnology and Biological Sciences Research Council http://dx.doi.org/10.13039/501100000268
                Award ID: BB/K01997X/1
                Award ID: BB/I00095X/1
                Award ID: BB/L024225/1
                Funded by: Wellcome Trust http://dx.doi.org/10.13039/100004440
                Award ID: WT101477MA
                Funded by: European Commission http://dx.doi.org/10.13039/501100000780
                Award ID: PRIME-XS [grant number 262067]
                Award ID: ProteomeXchange [grant number 260558]
                Funded by: Natural Science Foundation of the Hubei Province (China)
                Award ID: 2010CDB01401
                Funded by: Educational Commission of the Hubei Province
                Award ID: Q20113001
                Funded by: North Rhine-Westphalia, Germany
                Award ID: PURE (Protein Unit for Research in Europe)
                Categories
                Technological Innovation and Resources

                Molecular biology
                Molecular biology

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