Glycoproteomic Alterations in Drug-Resistant Nonsmall Cell Lung Cancer Cells Revealed by Lectin Magnetic Nanoprobe-Based Mass Spectrometry
datasetposted on 28.09.2018, 00:00 authored by Juanilita T. Waniwan, Yi-Ju Chen, Rey Capangpangan, Shao-Hsing Weng, Yu-Ju Chen
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 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.
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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