Rapid De-O-Glycosylation Concomitant with Peptide Labeling Using Microwave Radiation and an Alkyl Amine Base

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• 作者：Stephanie Maniatis ; Hui Zhou ; Vernon Reinhold
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：March 15, 2010
• 年：2010
• 卷：82
• 期：6
• 页码：2421-2425
• 全文大小：231K
• 年卷期：v.82,no.6(March 15, 2010)
• ISSN：1520-6882

文摘

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.