Title : Selective
glycopeptide profiling by acetone enrichment and LC/MS
Abstract :
- LC/MS is commonly used for site-specific glycosylation analysis of glycoproteins in cells and tissues
- A limitation of this technique is the difficulty in acquiring reliable mass spectra for glycopeptides , mainly due to their high heterogeneity and poor hydrophobicity
- Here, we establish a versatile method for efficient glycopeptide enrichment to acquire reliable mass spectra
- Several lines of evidence using model glycoproteins suggest that our method is based on the different solubility between non-glycosylated and glycosylated peptides in acetone
- We also provide data showing that the acetone-precipitated glycopeptide enrichment was successful in acquiring a more comprehensive MS/MS data set for the various glycoforms of each glycopeptide in crude human serum
- We propose that this method is a powerful tool for the acquisition of reliable mass spectra from trace amounts of glycopeptides and an alternative to lectin affinity enrichment
- BIOLOGICAL SIGNIFICANCE: In this study, we established a versatile method for glycopeptide enrichment to acquire reliable mass spectra because of the limitation of conventional enrichment methods, such as lectin affinity chromatography
- Our enrichment method is capable of isolating glycopeptides from complex peptide mixtures such as crude serum