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A Quantitative Glycomics and Proteomics Combined Purification Strategy
Published on: March 8, 2016
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Sialic acid linkage-specific quantitative N-glycoproteomics using selective alkylamidation and multiplex TMT-labeling
1School of Chemical Science & Engineering, Shanghai Key Laboratory of Chemical Assessment and Sustainability, Tongji University, Shanghai, 200092, China.
Analytica Chimica Acta
|October 3, 2022
Summary
This study introduces a new method for analyzing sialic acid linkage-specific N-glycoproteomics in intact N-glycopeptides. The approach identified differential sialylation in lung cancer, revealing potential biomarkers and links to COVID-19.
Area of Science:
- Biochemistry
- Proteomics
- Glycobiology
Background:
- Protein sialylation is crucial for biological processes, and its dysregulation is linked to diseases like cancer.
- Quantitative analysis of aberrant sialylation at the intact N-glycopeptide level is challenging due to a lack of multiplexing methods.
Purpose of the Study:
- To develop and apply a novel sialic acid linkage-specific quantitative N-glycoproteomics method.
- To characterize differential sialylation in lung cancer tissues compared to non-tumor tissues at the intact N-glycopeptide level.
Main Methods:
- Utilized selective alkylamidation and multiplex tandem mass tags (TMT)-labeling for quantitative N-glycoproteomics.
- Employed reversed-phase liquid chromatography-nano-electron spray ionization-tandem mass spectrometry (RPLC-nanoESI-MS/MS) for comprehensive analysis.
- Analyzed intact N-glycopeptides with and without sialic acid modifications.
Main Results:
- Identified 6384 intact N-glycopeptides and quantified 521 differentially expressed ones from 254 N-glycoproteins in lung cancer.
- Discovered 307 sialylated intact N-glycopeptides, including specific α2,6- and α2,3-linked sialic acids.
- Found associations between α2,6-sialylation and COVID-19, and characterized various N-glycosylation types in immunoglobulin family proteins.
Conclusions:
- The developed method enables linkage-specific quantitative analysis of sialylation at the intact N-glycopeptide level.
- Differential sialylation patterns in lung cancer were identified, highlighting potential biomarkers.
- The findings provide insights into the role of sialylation in cancer and infectious diseases.
