cancel
Showing results for 
Search instead for 
Did you mean: 
Orbitrap_SciLib
Reputable Mentor II
Reputable Mentor II
Darula Z, Sherman J, Medzihradszky KF.
Mol Cell Proteomics. 2012 Jul;11(7):O111.016774.
Two different workflows were tested in order to develop methods that provide deeper insight into the secreted O-glycoproteome. Bovine serum samples were subjected to lectin affinity-chromatography both at the protein- and peptide-level in order to selectively isolate glycopeptides with the most common, mucin core-1 sugar. This enrichment step was implemented with either protein-level mixed-bed ion-exchange chromatography or with peptide-level electrostatic repulsion hydrophilic interaction chromatography. Both methods led to at least 65% of the identified products being glycopeptides, in comparison to ∼25% without the additional chromatography steps [Darula, Z., and Medzihradszky, K. F. (2009) Affinity enrichment and characterization of mucin core-1 type glycopeptides from bovine serum. Mol. Cell. Proteomics 8, 2515-2526]. In order to improve not only the isolation but also the characterization of the glycopeptides exoglycosidases were used to eliminate carbohydrate extensions from the directly peptide-bound GalNAc units. Consequent tandem MS analysis of the mixtures using higher-energy collision-dissociation and electron-transfer dissociation led to the identification of 124 glycosylation sites in 51 proteins. While the electron-transfer dissociation data provided the bulk of the information for both modified sequence and modification site assignment, the higher-energy collision-dissociation data frequently yielded confirmation of the peptide identity, and revealed the presence of some core-2 or core-3 oligosaccharides. More than two-thirds of the sites as well as the proteins have never been reported modified.

http://www.mcponline.org/content/11/7/O111.016774.abstract
Proteomics Research Group, Biological Research Center of the Hungarian Academy of Sciences, Szeged, H-6701, Szeged, POB 521, Hungary;
Version history
Last update:
‎10-15-2021 11:09 AM
Updated by:
Closed Account
Contributors