Simultaneous and sialic acid linkage-specific N- and O-linked glycan analysis by ester-to-amide derivatization.

Autor: Hanamatsu H; Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Hokkaido, 060-8638, Sapporo, Japan. h_hanamatsu@med.hokudai.ac.jp.; Department of Gastroenterology and Hepatology, Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido, 060-8638, Japan. h_hanamatsu@med.hokudai.ac.jp., Miura Y; Sumitomo Bakelite Co., Ltd., 5-8, Tennoz Parkside Building, Higashi-Shinagawa 2-chome, Shinagawa-ku, 140-0002, Tokyo, Japan., Nishikaze T; Solutions COE, Analytical & Measuring Instruments Division, Shimadzu Corporation, 604-8511, Kyoto, Japan., Yokota I; Institute for Glyco-core Research (iGCORE), Nagoya University, 464-8601, Nagoya, Japan., Homan K; Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Hokkaido, 060-8638, Sapporo, Japan., Onodera T; Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Hokkaido, 060-8638, Sapporo, Japan., Hayakawa Y; Solutions COE, Analytical & Measuring Instruments Division, Shimadzu Corporation, 604-8511, Kyoto, Japan., Iwasaki N; Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Hokkaido, 060-8638, Sapporo, Japan., Furukawa JI; Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Hokkaido, 060-8638, Sapporo, Japan. j-furu@med.nagoya-u.ac.jp.; Institute for Glyco-core Research (iGCORE), Nagoya University, 464-8601, Nagoya, Japan. j-furu@med.nagoya-u.ac.jp.
Jazyk: angličtina
Zdroj: Glycoconjugate journal [Glycoconj J] 2023 Apr; Vol. 40 (2), pp. 259-267. Date of Electronic Publication: 2023 Mar 06.
DOI: 10.1007/s10719-023-10109-8
Abstrakt: Characterization of O-glycans linked to serine or threonine residues in glycoproteins has mostly been achieved using chemical reaction approaches because there are no known O-glycan-specific endoglycosidases. Most O-glycans are modified with sialic acid residues at the non-reducing termini through various linkages. In this study, we developed a novel approach for sialic acid linkage-specific O-linked glycan analysis through lactone-driven ester-to-amide derivatization combined with non-reductive β-elimination in the presence of hydroxylamine. O-glycans released by non-reductive β-elimination were efficiently purified using glycoblotting via chemoselective ligation between carbohydrates and a hydrazide-functionalized polymer, followed by modification of methyl or ethyl ester groups of sialic acid residues on solid-phase. In-solution lactone-driven ester-to-amide derivatization of ethyl-esterified O-glycans was performed, and the resulting sialylated glycan isomers were discriminated by mass spectrometry. In combination with PNGase F digestion, we carried out simultaneous, quantitative, and sialic acid linkage-specific N- and O-linked glycan analyses of a model glycoprotein and human cartilage tissue. This novel glycomic approach will facilitate detailed characterization of biologically relevant sialylated N- and O-glycans on glycoproteins.
(© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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