Journal
ANALYTICAL CHEMISTRY
Volume 82, Issue 6, Pages 2421-2425Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac902734w
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Funding
- NIH
- NIGMS [GM 54045]
- Glycan Connections, Lee, NH
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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 MSn 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 with both N- and O-linked glycans and a highly glycosylated swine mucin.
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