title: Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues.
abstract: 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 β-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.
authors.affiliation: †Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Furukawa
authors.affiliation: †Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Piao
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Yoshida
authors.affiliation: †Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Kazue
authors.initials: K
authors.lastname: Okada
authors.affiliation: †Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Ikuko
authors.initials: I
authors.lastname: Yokota
authors.forename: Kenichi
authors.initials: K
authors.lastname: Higashino
authors.affiliation: §Graduate School of Environmental Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Sakairi
authors.affiliation: †Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Yasuro
authors.initials: Y
authors.lastname: Shinohara
mesh_terms: D000818:Animals; D054794:Glycomics / Q000379:methods*; D006031:Glycosylation; D008099:Liver / Q000737:chemistry; D051379:Mice; D008872:Microwaves; D011134:Polysaccharides / Q000737:chemistry*; D047069:Pyrazolones / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D011134:Polysaccharides; D047069:Pyrazolones; C038362:pyrazolone