Quantitative O-Glycomics by Microwave-Assisted 尾-Elimination in the Presence of Pyrazolone Analogues

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• 作者：Jun-ichi Furukawa ; Jinhua Piao ; Yasunobu Yoshida ; Kazue Okada ; Ikuko Yokota ; Kenichi Higashino ; Nobuo Sakairi ; Yasuro Shinohara
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：August 4, 2015
• 年：2015
• 卷：87
• 期：15
• 页码：7524-7528
• 全文大小：321K
• ISSN：1520-6882

文摘

O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. Therefore, chemical digestion approaches have been widely used; however, it is sometimes difficult to suppress unwanted side reactions. Recently, we reported a novel O-glycomics procedure using 尾-elimination in the presence of pyrazolone analogues (BEP). In the present study, we describe a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis. Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples.