Detergent-Assisted Glycoprotein Capture: A Versatile Tool for In-Depth N-Glycoproteome Analysis

J Proteome Res. 2016 Jun 3;15(6):2080-6. doi: 10.1021/acs.jproteome.6b00056. Epub 2016 May 12.


Large-scale N-glycoproteome studies have been hindered by poor solubility of hydrophobic membrane proteins and the complexity of proteome samples. Herein, we developed a detergent-assisted glycoprotein capture method to reduce these issues by conducting hydrazide chemistry-based glycoprotein capture in the presence of strong detergents such as sodium dodecyl sulfate and Triton X-100. The strong detergents helped to solubilize hydrophobic membrane proteins and then increased the access of hydrazide groups to oxidized glycoproteins, thus increasing the coverage of the N-glycoproteome. Compared with the conventional glycopeptide capture method, the detergent-assisted glycoprotein capture approach nearly doubled the number of N-glycosylation sites identified from HEK 293T cells with improved specificity. Application of this approach in the larger scale N-glycoproteomics analysis of the HEK 293T cell membrane led to the identification of 2253 unique N-glycosites from 953 proteins. Furthermore, the application of this approach to human serum resulted in the identification of 850 N-glycosylation sites without any immunodepletion or fractionation. Overall, the detergent-assisted glycoprotein capture method simplified the capture process, and it increased the number of sites observed on both hydrophobic membrane proteins and hydrophilic secreted proteins.

Keywords: N-glycoproteome; detergent; glycoprotein capture; membrane; serum.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Detergents / chemistry*
  • Glycoproteins / analysis*
  • HEK293 Cells
  • Humans
  • Membrane Proteins / analysis*
  • Proteome / analysis
  • Proteomics / methods*
  • Serum / chemistry


  • Detergents
  • Glycoproteins
  • Membrane Proteins
  • Proteome