PXD061371

PXD061371 is an original dataset announced via ProteomeXchange.

Title	High-Throughput Simultaneous Quantification of Glycopeptides and Phosphopeptides Enabled by 12-plex DiLeu Isobaric Tags and Dual-Functional Titanium(IV)-IMAC Material
Description	Protein glycosylation and phosphorylation are two of the most common post-translational modifications (PTMs). Though they have been indicated to play critical roles in various physiological and pathological processes, effective analytical tools are challenged by low PTM abundance and poor ionization efficiency. Recently, an epoxy-ATP-Ti4+-IMAC material has been developed to enable simultaneous enrichment and separation of common N-glycopeptides, phosphopeptides, and mannose-6-phosphate (M6P) glycopeptides from complex biological samples. To further improve the throughput of our previous method, in this study, we developed an efficient strategy using our custom-developed N,N-dimethyl leucine (DiLeu) isobaric tags to achieve a multiplexed quantitative simultaneous analysis of N-glycosylation and phosphorylation from up to 12 samples for the first time. We further applied this method to investigate N-glycosylation and phosphorylation alterations in APP/PS1 model mouse brain samples and wild-type (WT) controls. With the integration of a one-tube sample processing workflow, 1975 glycopeptides and 1181 phosphopeptides were quantitatively identified from mouse brain. We also unveiled important biological functions regulated by N-glycosylation and phosphorylation, including synapse organization, synaptic membrane and cell adhesion, highlighting the involvement of N-glycosylation and phosphorylation aberrations in APP/PS1 models. Our results provide evidence that the established DiLeu isobaric labeling-epoxy-Ti4+-IMAC method can be used for high-throughput simultaneous quantification of glycosylation and phosphorylation, and their potential crosstalk, showing significant promise for identifying novel therapeutic targets or biomarkers in biological systems.
HostingRepository	PRIDE
AnnounceDate	2026-06-29
AnnouncementXML	Submission_2026-06-28_16:39:16.964.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Feixuan Wu
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606; scientific name: Mus musculus (Mouse); NCBI TaxID: NEWT:10090;
ModificationList	N-glycosylated residue; phosphorylated residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2025-03-02 18:14:24	ID requested
⏵ 1	2026-06-28 16:39:17	announced

10.1016/j.aca.2026.345231;

Wu F, Wang D, Tabang DN, Liu PK, Wang Z, Liu Y, Steenhagen A, Puglielli L, Li L, High-throughput simultaneous quantification of glycopeptides and phosphopeptides enabled by 12-plex DiLeu isobaric tags and dual-functional Titanium(IV)-IMAC material. Anal Chim Acta, 1395():345231(2026) [pubmed]

submitter keyword: Phosphoproteomics,Glycoproteomics, Enrichment, Post-translational modifications, High-throughput quantitation, Immobilized metal affinity chromatography

Lingjun Li
contact affiliation	School of Pharmacy, Department of Chemistry and Biophysics Graduate Program, University of Wisconsin-Madison, Madison, Wisconsin 53705-2222, USA
contact email	lingjun.li@wisc.edu
lab head
Feixuan Wu
contact affiliation	University of Wisconsin-Madison
contact email	wufeixuan07@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD061371
     1. Label: PRIDE project
     2. Name: High-Throughput Simultaneous Quantification of Glycopeptides and Phosphopeptides Enabled by 12-plex DiLeu Isobaric Tags and Dual-Functional Titanium(IV)-IMAC Material