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

Significance Statement

Quantification of O-glycomics found on the cell surface is important in physiology and pathology. Quantification will aid in understanding role of glycans in bacterial and viral recognition, cellular signaling pathways, innate immunity, cancer development. Additionally there are many glycan-specific diseases and advancing glycomic technologies will provide better diagnosis and therapy. This study describe an improved method in quantification of o-glycomics using pyrazolone analogues.

Journal Reference

Anal Chem. 2015;87(15):7524-8.

Furukawa J1, Piao J1, Yoshida Y, Okada K1, Yokota I1, Higashino K, Sakairi N2, Shinohara Y1. 

Show Affiliations
  1. Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Hokkaido University, Sapporo 001-0021, Japan.
  2. Graduate School of Environmental Science, Hokkaido University, Sapporo 060-0810, Japan.

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 pyrazolone analogues  procedure for rapid and quantitative O-glycomic analysis. Following optimization of the reaction conditions, the MW-assisted pyrazolone analogues  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 pyrazolone analogues  procedure enabled O-glycomic analyses of various biological samples.

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Quantitative O-Glycomics by Microwave-Assisted β-Elimination Presence of Pyrazolone Analogues. Global Medical Discovery feature