PXD061247

PXD061247 is an original dataset announced via ProteomeXchange.

Title	Integrated Single-Tip IMAC-HILIC Enrichment Enables Simultaneous and Efficient Analysis of Plant Phosphoproteomics and N-glycoproteomics
Description	Protein phosphorylation and N-glycosylation are key post-translational modifications (PTMs) in plants that regulate critical signaling processes. However, analyzing both PTMs simultaneously remains challenging due to low abundance and divergent enrichment requirements. Here, we present a single-tip IMAC-HILIC approach that integrates immobilized metal affinity chromatography (IMAC) and hydrophilic interaction chromatography (HILIC) materials within one pipette tip, enabling concurrent enrichment and sequential elution of phosphopeptides and N-glycopeptides. This workflow effectively reduces coelution of phosphopeptides during N-glycopeptide elution and significantly streamlines sample processing. Benchmarking against the tandem-tip TIMAHAC method demonstrates comparable identification depth and superior quantitative accuracy for N-glycopeptides. We further apply IMAC-HILIC to examine calcium deprivation in Arabidopsis, revealing distinct global changes in both phosphoproteome and N-glycoproteome. Our optimized protocol offers a practical, high-throughput solution for dual PTM profiling in complex plant samples, paving the way for broader investigations for PTM crosstalk in diverse physiological and stress responses.
HostingRepository	PRIDE
AnnounceDate	2025-05-14
AnnouncementXML	Submission_2025-05-13_19:19:41.599.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ting-An Chen
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	N-glycosylated residue; phosphorylated residue
Instrument	timsTOF HT

Revision	Datetime	Status	ChangeLog Entry
0	2025-02-26 21:14:44	ID requested
⏵ 1	2025-05-13 19:19:41	announced

Dataset with its publication pending

submitter keyword: Phosphoproteomics

N-glycoproteomics

Enrichment

Immobilized Metal Affinity Chromatography

Hydrophilic interaction Chromatography

Chuan-Chih Hsu
contact affiliation	Institute of Plant and Microbial Biology, Academia Sinica
contact email	cchsu@gate.sinica.edu.tw
lab head
Ting-An Chen
contact affiliation	IPMB, Academia Sinica
contact email	annie1850331@gmail.com
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/2025/05/PXD061247

PRIDE project URI

• PRIDE
  1. PXD061247
     1. Label: PRIDE project
     2. Name: Integrated Single-Tip IMAC-HILIC Enrichment Enables Simultaneous and Efficient Analysis of Plant Phosphoproteomics and N-glycoproteomics