PXD062515 is an
original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Scalable Phosphotyrosine Enrichment with SH2 Superbinder Enables Deep Profiling of EGF Responses |
| Description | Phosphotyrosine signaling plays a critical role in many biological processes, from cell proliferation to immune response. Despite its importance, proteomic studies of tyrosine phosphorylation have been limited in scale and throughput by costly reagents and labor-intensive protocols. To address this, we developed R2HaPpY, a phosphotyrosine enrichment method combining simplified superbinder reagent preparation and automated high-throughput enrichment. Our new reagent binds phosphotyrosine peptides at higher efficiency than existing enrichment reagents and reduces both cost and preparation time by 20-fold. We generalized R2HaPpY to samples of low and high phosphotyrosine levels, and benchmarked it on EGF signaling dynamics in HeLa cells. Using only ~1 mg of input peptides, we quantify 1,651 unique phosphotyrosine sites, including 878 regulated phosphotyrosine sites, many novel or not previously annotated as EGF-responsive. Our results reveal differential temporal regulation and represent the largest phosphotyrosine dataset of EGF stimulation response to date. This streamlined, cost-effective, and sensitive method enables quantitative mapping of tyrosine phosphorylation dynamics at the scale of hundreds of samples, facilitating integration of phosphotyrosine signaling into multiomic studies across diverse biological systems. |
| HostingRepository | PRIDE |
| AnnounceDate | 2026-07-03 |
| AnnouncementXML | Submission_2026-07-03_13:23:36.737.xml |
| DigitalObjectIdentifier | https://doi.org/10.6019/PXD062515 |
| ReviewLevel | Peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Supported dataset by repository |
| PrimarySubmitter | Alexis Chang |
| SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: NEWT:4932; scientific name: Saccharomyces cerevisiae; NCBI TaxID: NCBITaxon:4932; |
| ModificationList | phosphorylated residue |
| Instrument | Orbitrap Exploris 480 |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|
| 0 | 2025-04-02 04:14:36 | ID requested | |
| ⏵ 1 | 2026-07-03 13:23:37 | announced | |
Publication List
Keyword List
| submitter keyword: phosphotyrosine, automated, HeLa, enrichment, high-throughput, EGF, LC-MS/MS |
Contact List
| Judit Villen |
|---|
| contact affiliation | Department of Genome Sciences; Villen Lab; University of Washington; United States |
| contact email | jvillen@uw.edu |
| lab head | |
| Alexis Chang |
|---|
| contact affiliation | University of Washington |
| contact email | alexisn1@uw.edu |
| dataset submitter | |
Full Dataset Link List
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/2026/07/PXD062515 |
| PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD062515
- Label: PRIDE project
- Name: Scalable Phosphotyrosine Enrichment with SH2 Superbinder Enables Deep Profiling of EGF Responses
