Glycoproteomic
Alterations in Drug-Resistant Nonsmall
Cell Lung Cancer Cells Revealed by Lectin Magnetic Nanoprobe-Based
Mass Spectrometry
Juanilita
T. Waniwan
Yi-Ju Chen
Rey Capangpangan
Shao-Hsing Weng
Yu-Ju Chen
10.1021/acs.jproteome.8b00433.s002
https://acs.figshare.com/articles/dataset/Glycoproteomic_Alterations_in_Drug-Resistant_Nonsmall_Cell_Lung_Cancer_Cells_Revealed_by_Lectin_Magnetic_Nanoprobe-Based_Mass_Spectrometry/7218794
Understanding 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 <i>N-</i>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.
2018-09-28 00:00:00
site-specific function
sialylated glycopeptides
EGFR-TKI-sensitive PC 9
glycosylation-mediated pathogenesis
Glycoproteomic Alterations
drug resistance
site-specific glycosylation analysis
NSCLC resistance
PC 9-IR cells
MNP
glycoproteome
HCD-CID-MS
Nanoprobe-Based Mass Spectrometry
nonsmall cell lung cancer
onco-protein EGFR
Label-free quantitation
glycoproteomic atlas
51 glycopeptides
Drug-Resistant Nonsmall Cell Lung Cancer Cells Revealed
2767 nonredundant glycopeptides
multilectin nanoprobe
glycotope-specific enrichment