PXD029269-2

PXD029269 is an original dataset announced via ProteomeXchange.

Title	Boost-DiLeu: Enhanced Isobaric N,N-Dimethyl Leucine Tagging Strategy for Comprehensive Quantitative Glycoproteomic Analysis
Description	Intact glycopeptide analysis has been of great interest because it can elucidate glycosylation site information and glycan structural composition at the same time. However, mass-spectrometry (MS)-based glycoproteomic analysis is hindered by the low stoichiometry of glycosylation and poor ionization efficiency of glycopeptides. Due to the relatively large amounts of starting materials needed for enrichment, identification and quantification of intact glycopeptides in some size-limited biological systems are especially challenging. To overcome these limitations, here we developed an improved boosting strategy to enhance N,N-dimethyl leucine tagging-based quantitative glycoproteomic analysis, termed as Boost-DiLeu. With integration of one-tube sample processing workflow and high pH fractionation, 3514 quantifiable N-glycopeptides were identified from 30 µg HeLa cell tryptic digests with reliable quantification performance. Furthermore, this strategy was applied to human cerebrospinal fluid (CSF) samples to differentiate N-glycosylation profiles between Alzheimer’s disease patients and healthy donors. The results revealed processes and pathways affected by dysregulated N-glycosylation in AD, including platelet degranulation, cell adhesion, and extracellular matrix, which highlighted the involvement of N-glycosylation aberrations in AD pathogenesis. Moreover, co-expression network analysis (WGCNA) showed 3 modules of glycoproteins, one of which are associated with AD phenotype. Our results demonstrated the feasibility of using this strategy for comprehensive glycoproteomic analysis of size-limited clinical samples. Taken together, we developed and optimized a strategy for enhanced comprehensive quantitative intact glycopeptide analysis with DiLeu labeling, which is especially promising for identifying novel therapeutic targets or biomarkers in sample size limited models or systems.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:53:42.345.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Danqing Wang
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	complex glycosylation; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2021-10-21 05:10:22	ID requested
1	2022-09-07 17:47:47	announced
⏵ 2	2023-11-14 08:53:42	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: Human, Glycoproteomics, CSF, LC-MS/MS

Lingjun Li
contact affiliation	University of Wisconsin-Madison, Department of Chemistry and School of Pharmacy
contact email	lingjun.li@wisc.edu
lab head
Danqing Wang
contact affiliation	University of Wisconsin-Madison
contact email	dwang356@wisc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD029269
     1. Label: PRIDE project
     2. Name: Boost-DiLeu: Enhanced Isobaric N,N-Dimethyl Leucine Tagging Strategy for Comprehensive Quantitative Glycoproteomic Analysis