PXD007556

PXD007556 is an original dataset announced via ProteomeXchange.

Title	Protease specificity profiling in a pipette tip using “charge-synchronized” proteome-derived peptide libraries
Description	About 2% of the genome of human and other organisms codes for proteases. An important step toward deciphering the biological function of a protease and designing inhibitors is the profiling of protease specificity. In this work we present a novel, label-free, proteomics-based protease specificity profiling method that only requires simple sample preparation steps. It uses proteome-derived peptide libraries and enriches the cleaved sequences using strong cation exchange chromatography (SCX) material in a pipette tip. As a demonstration of the method’s versatility, we successfully determined the specificity of GluC, caspase-3, chymotrypsin and cathepsin G from several hundreds to almost 2000 cleavage events per proteases. Interestingly, we also found and confirmed a novel intrinsic preference of cathepsin G for Asn at the P1 subsite, explaining the reported cleavage position on the P. aeruginosa aeruginosa flagellin by human cathepsin G . Overall, this method is straightforward and requires so far the lowest investment in material and equipment for protease specificity profiling. Therefore, we think it will be applicable in any biochemistry laboratory and promote an increased understanding of protease specificity.
HostingRepository	PRIDE
AnnounceDate	2018-04-26
AnnouncementXML	Submission_2018-04-26_05:56:53.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Minh Thi Nhat Nguyen
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	acetylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-08-28 00:57:20	ID requested
⏵ 1	2018-04-26 05:56:54	announced

Nguyen MTN, Shema G, Zahedi RP, Verhelst SHL, Protease Specificity Profiling in a Pipet Tip Using "Charge-Synchronized" Proteome-Derived Peptide Libraries. J Proteome Res, 17(5):1923-1933(2018) [pubmed]

curator keyword: Technical

submitter keyword: ChaFRAtip, protease specificity, label-free, charged-synchronized peptide library

Steven L. Verhelst
contact affiliation	Prof. Dr. Steven Verhelst Laboratory of Chemical Biology Department of Cellular and Molecular Medicine KU Leuven - University of Leuven Herestraat 49, box 802 3000 Leuven Belgium
contact email	stevenverhelst@kuleuven.de
lab head
Minh Thi Nhat Nguyen
contact affiliation	Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V.
contact email	minh.nguyen@isas.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/04/PXD007556

PRIDE project URI

• PRIDE
  1. PXD007556
     1. Label: PRIDE project
     2. Name: Protease specificity profiling in a pipette tip using “charge-synchronized” proteome-derived peptide libraries