Rapid de-O-glycosylation concomitant with peptide labeling using microwave radiation and an alkyl amine base.

ABSTRACT: Procedures are detailed for a quantitative release of O-linked glycans from peptides that now provide a shorter reaction time, a possible identification of O-linked sites, and a quantification of all reaction products. The release was initiated by a mild base, dimethylamine, and accelerated by microwave radiation. Differential analysis using standard glycoproteins has shown improved release efficiency concurrent with facile incorporation of dimethylamine into the former O-linked sites. In situ glycan reduction insures protection against peeling and is synchronous with subsequent studies by high performance MS(n) sequencing. The protocols were established with a synthetic O-GlcNAc peptide that would mimic the linkage chemistry and applied to a well characterized glycoprotein bovine fetuin w

SUBMITTER: Maniatis S 

PROVIDER: S-EPMC2837759 | biostudies-literature | 2010 Mar

REPOSITORIES: biostudies-literature