PXD043713-2

PXD043713 is an original dataset announced via ProteomeXchange.

Title	A bespoke analytical workflow for the identification of sulfopep-tides and confident discrimination from phosphopeptides_C4PR_LIV
Description	Protein tyrosine sulfation (sY) is a post-translational modification (PTM) catalysed by Golgi-resident Tyrosyl Protein Sul-foTransferases (TPSTs). Information on protein tyrosine sulfation is currently limited to ~50 human proteins with only a handful of those having verified sites of sulfation. The contribution of this chemical moiety for the regulation of biological processes, both inside and outside the cell, remains poorly defined in large part due to analytical limitations. Mass spectrom-etry-based proteomics is the method of choice for PTM analysis, but has yet to be applied for the systematic investigation of biological sulfation (the ‘sulfome’), primarily due to issues associated with discrimination of sY- from phosphotyrosine (pY)-containing peptides. In this study, we developed a mass spectrometry (MS)-based workflow centred on the characteri-zation of sY-peptides, incorporating optimised Zr4+-IMAC and TiO2 enrichment strategies. Extensive characterization of a panel of sY- and pY-peptides using an array of MS fragmentation regimes (CID, HCD, EThcC, ETciD, UVPD) highlights differences in the ability to generate site-determining product ions, which can be exploited to differentiate sulfated peptides from nominally isobaric phosphopeptides based on precursor ion neutral loss at low collision energy. Application of our ana-lytical workflow to the HEK-293 cell extracellular secretome facilitated identification of 23 new sulfotyrosine-containing peptides, several of which we validate enzymatically using in vitro sulfation assays. Our study demonstrates the applicabil-ity of this strategy for confident, high-throughput, ‘sulfomics’ studies
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_06:27:33.672.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	leonard daly
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	sulfated residue; phosphorylated residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-07-12 05:19:02	ID requested
1	2024-01-26 10:36:32	announced
⏵ 2	2024-10-22 06:27:34	announced	2024-10-22: Updated project metadata.

Daly LA, Byrne DP, Perkins S, Brownridge PJ, McDonnell E, Jones AR, Eyers PA, Eyers CE, Custom Workflow for the Confident Identification of Sulfotyrosine-Containing Peptides and Their Discrimination from Phosphopeptides. J Proteome Res, 22(12):3754-3772(2023) [pubmed]

10.1021/acs.jproteome.3c00425;

submitter keyword: tyrosine-o-sulfation, sulfotyrosine,sulfation

Claire eyers
contact affiliation	Centre for Proteome Research, Institute of Systems, Molecular & Integrative Biology, University of Liverpool, Crown Street, Liverpool, L69 7ZB, U.K
contact email	ceyers@liverpool.ac.uk
lab head
leonard daly
contact affiliation	University of Liverpool
contact email	leonard.daly@liverpool.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD043713
     1. Label: PRIDE project
     2. Name: A bespoke analytical workflow for the identification of sulfopep-tides and confident discrimination from phosphopeptides_C4PR_LIV