Related Experiment Video
Updated: May 18, 2026
10:11
Glycopeptide Capture for Cell Surface Proteomics
Published on: May 9, 2014
A new method for quantitative analysis of cell surface glycoproteome
Zhen Sun1, Rui Chen, Kai Cheng
1Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
Proteomics
|September 25, 2012
Summary
This study introduces a novel glycoproteomics method for precise cell surface glycosylation analysis. The approach accurately quantifies glycoproteins, distinguishing changes in protein abundance from glycosite occupancy, aiding biomarker discovery.
Area of Science:
- Biochemistry
- Proteomics
- Cell Biology
Background:
- Altered cell surface protein glycosylation is linked to various diseases.
- Traditional glycoproteomics methods struggle to differentiate protein abundance changes from glycosite occupancy variations.
- Accurate quantification of cell surface glycosylation is crucial for understanding disease mechanisms.
Purpose of the Study:
- To develop and validate a modified cell surface-capturing strategy for accurate and specific quantification of cell surface glycosylation profiles.
- To differentiate changes in protein abundance from alterations in glycosite occupancy.
- To identify differentially expressed cell surface glycoproteins between two human cell lines.
Main Methods:
- A modified cell surface-capturing strategy was employed.
- Glycopeptides were directly analyzed using LC-MS/MS for glycoprotein identification.
- Non-glycopeptides were isotopically labeled for glycoprotein quantification.
Main Results:
- The strategy identified 341 glycoproteins with 82.4% specificity for cell membrane proteins.
- 33 glycoproteins exhibited significant expression changes between Chang Liver and HepG2 cells.
- Differential expression of EMMPRIN and BCAM was confirmed by Western blotting.
Conclusions:
- The developed method allows for specific and accurate analysis of cell surface glycoproteins.
- This approach effectively distinguishes protein abundance from glycosite occupancy changes.
- The method holds potential for broad applications in biomarker and drug target discovery.
