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PXD002785

PXD002785 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleTag-Free Identification of Protease Specificity Using Proteome-Derived Peptide Libraries
DescriptionProteome-derived peptide libraries are a powerful tool for the investigation of protease specificity, conventionally involving the biotinylation of cleavage products to enable for their enrichment. Here we present a modified strategy for protease specificity profiling using proteome-derived peptide libraries in a tag-free manner. In comparison to the original method the new protocol is faster, avoids cost- and time-intensive enrichment steps, and enables probing for lysine selectivity. In the new workflow, peptide libraries are generated by endoproteolytic digestion of proteomes without chemical modification of primary amines prior to exposure to a protease under investigation. After incubation with the test protease, treated and control library are differentially labeled using stable isotopes by means of reductive dimethylation. Upon analysis by liquid chromatography mass spectrometry, cleavage products of the test protease appear as peptides enriched for the respective isotopic label. With “PICS 2.0” we identified more than 2800 cleavage sites in five specificity experiments for four proteases, including trypsin, caspase-3, and chlamydial protease-like activity factor (CPAF). Hence, this method is a valuable addition to the different strategies currently employed for specificity profiling of proteases
HostingRepositoryPRIDE
AnnounceDate2016-05-02
AnnouncementXMLSubmission_2016-05-02_00:42:58.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterOliver Schilling
SpeciesList scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationListcarbamoylated residue; dimethylated residue
InstrumentQ Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02015-08-24 06:56:19ID requested
12016-05-02 00:43:00announced
Publication List
Biniossek ML, Niemer M, Maksimchuk K, Mayer B, Fuchs J, Huesgen PF, McCafferty DG, Turk B, Fritz G, Mayer J, Haecker G, Mach L, Schilling O, Identification of Protease Specificity by Combining Proteome-Derived Peptide Libraries and Quantitative Proteomics. Mol Cell Proteomics, 15(7):2515-24(2016) [pubmed]
Keyword List
curator keyword: Technical
submitter keyword: Proteolysis
Contact List
Oliver Schilling
contact affiliationUniversity of Freiburg
contact emailoliver.schilling@mol-med.uni-freiburg.de
lab head
Oliver Schilling
contact affiliationUniversity of Freiburg
contact emailoliver.schilling@mol-med.uni-freiburg.de
dataset submitter
Full Dataset Link List
Dataset FTP location
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PRIDE project URI
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