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PXD025536

PXD025536 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleNano-scale solid-phase isobaric labeling for multiplexed quantitative phosphoproteomics
DescriptionWe established a workflow for highly sensitive multiplexed quantitative phosphoproteomics using a nano-scale solid-phase tandem mass tag (TMT) labeling reactor. Phosphopeptides were firstly enriched by titanium oxide chromatography and then labeled with isobaric TMT reagents in a StageTip packed with hydrophobic polymer-based sorbents. We found that TMT-labeled singly phosphorylated peptides tend to flow through the titanium oxide column. Therefore, TMT labeling should be performed after the enrichment step from tryptic peptides, resulting in the need for micro-scale reaction with small amounts of phosphopeptides. Using an optimized protocol for tens to hundreds of nanograms of phosphopeptides, we obtained a nearly 10-fold increase in sensitivity compared to the conventional solution-based TMT protocol. We demonstrate that this nano-scale phosphoproteomics protocol works for 50 μg of HeLa proteins treated with selumetinib, and we successfully quantified the selumetinib-regulated phosphorylated sites on a proteome scale. The MS raw data files have been deposited with the ProteomeXchange Consortium via the jPOST partner repository (http:// jpostdb.org) with the data set identifier PXD025536.
HostingRepositoryjPOST
AnnounceDate2021-07-23
AnnouncementXMLSubmission_2022-09-18_03:38:52.026.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterKosuke Ogata
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListS-carboxamidomethyl-L-cysteine; O-phospho-L-serine; TMT6plex reporter+balance reagent N-acylated residue; TMT6plex reporter+balance reagent acylated N-terminal; O-phospho-L-threonine; O4'-phospho-L-tyrosine; L-methionine sulfoxide; TMT; TMT; unknown modification; unknown modification
InstrumentQ Exactive; instrument
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-04-21 08:07:20ID requested
12021-07-23 05:55:41announced
22022-09-18 03:38:52announced2022-09-18: Updated FTP location.
Keyword List
submitter keyword: TMT labeling, solid-phase reactor, phosphopeptides, titanium oxide chromatography, multiplexed phosphoproteomics
Contact List
Yasushi Ishihama
lab head
Kosuke Ogata
contact affiliationKyoto university
dataset submitter
Full Dataset Link List
jPOST dataset URI
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.jpostdb.org/JPST001131/