pr8b00433_si_002.xlsx (3.81 MB)
Glycoproteomic Alterations in Drug-Resistant Nonsmall Cell Lung Cancer Cells Revealed by Lectin Magnetic Nanoprobe-Based Mass Spectrometry
dataset
posted on 2018-09-28, 00:00 authored by Juanilita
T. Waniwan, Yi-Ju Chen, Rey Capangpangan, Shao-Hsing Weng, Yu-Ju ChenUnderstanding the functional role
of glycosylation-mediated pathogenesis
requires deep characterization of glycoproteome, which remains extremely
challenging due to the inherently complex nature of glycoproteins.
We demonstrate the utility of lectin–magnetic nanoprobe (MNP@lectin)
coupled to Orbitrap HCD-CID-MS/MS for complementary glycotope-specific
enrichment and site-specific glycosylation analysis of the glycoproteome.
By three nanoprobes, MNP@ConA, MNP@AAL, and MNP@SNA, our results revealed
the first large-scale glycoproteome of nonsmall cell lung cancer (NSCLC)
with 2290 and 2767 nonredundant glycopeptides confidently identified
(Byonic score ≥100) in EGFR-TKI-sensitive PC9 and -resistant
PC9-IR cells, respectively, especially with more fucosylated and sialylated
glycopeptides in PC9-IR cells. The complementary enrichment was demonstrated
with only five glycopeptides commonly enriched in three MNP@lectins.
Glycotope specificity of 79 and 62% for enrichment was achieved using
MNP@AAL and MNP@SNA, respectively. Label-free quantitation revealed
predominant fucosylation in PC9-IR cells, suggesting its potential
role associated with NSCLC resistance. Moreover, without immunoprecipitation,
this multilectin nanoprobe allows the sensitive identification of
51 glycopeptides from 10 of 12 reported sites from onco-protein EGFR.
Our results not only demonstrated a sensitive approach to study the
vastly under-represented N-glycoprotome but also
may pave the way for a glycoproteomic atlas to further explore the
site-specific function of glycoproteins associated with drug resistance
in NSCLC.
History
Usage metrics
Categories
Keywords
site-specific functionsialylated glycopeptidesEGFR-TKI-sensitive PC 9glycosylation-mediated pathogenesisGlycoproteomic Alterationsdrug resistancesite-specific glycosylation analysisNSCLC resistancePC 9-IR cellsMNPglycoproteomeHCD-CID-MSNanoprobe-Based Mass Spectrometrynonsmall cell lung canceronco-protein EGFRLabel-free quantitationglycoproteomic atlas51 glycopeptidesDrug-Resistant Nonsmall Cell Lung Cancer Cells Revealed2767 nonredundant glycopeptidesmultilectin nanoprobeglycotope-specific enrichment
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC