Direct derivatization of sialic acids and mild β-elimination for linkage-specific sialyl O-glycan analysis.

Autor: Hanamatsu H; Institute for Glyco-core Research (iGCORE), Nagoya University, Nagoya, 464-8601, Japan., Yokota I; Institute for Glyco-core Research (iGCORE), Nagoya University, Nagoya, 464-8601, Japan., Kurogochi M; Institute for Glyco-core Research (iGCORE), Nagoya University, Nagoya, 464-8601, Japan; Laboratory of Glyco-Organic Chemistry, The Noguchi Institute, Tokyo, 173-0003, Japan., Akasaka-Manya K; Molecular Glycobiology, Research Team for Mechanism of Aging, Tokyo Metropolitan Institute for Geriatrics and Gerontology, Tokyo, 173-0015, Japan., Miura N; Institute for Glyco-core Research (iGCORE), Nagoya University, Nagoya, 464-8601, Japan., Manya H; Molecular Glycobiology, Research Team for Mechanism of Aging, Tokyo Metropolitan Institute for Geriatrics and Gerontology, Tokyo, 173-0015, Japan., Endo T; Molecular Glycobiology, Research Team for Mechanism of Aging, Tokyo Metropolitan Institute for Geriatrics and Gerontology, Tokyo, 173-0015, Japan., Nishikaze T; Solutions COE, Analytical & Measuring Instruments Division, Shimadzu Corporation, Kyoto, 604-8511, Japan. Electronic address: nishikaz@shimadzu.co.jp., Furukawa JI; Institute for Glyco-core Research (iGCORE), Nagoya University, Nagoya, 464-8601, Japan; Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, 060-8638, Japan. Electronic address: furukawa.junichi.n0@f.mail.nagoya-u.ac.jp., Tanaka K; Koichi Tanaka Mass Spectrometry Research Laboratory, Shimadzu Corporation, Kyoto, 604-8511, Japan.
Jazyk: angličtina
Zdroj: Analytica chimica acta [Anal Chim Acta] 2024 Aug 22; Vol. 1318, pp. 342945. Date of Electronic Publication: 2024 Jul 05.
DOI: 10.1016/j.aca.2024.342945
Abstrakt: Background: In sharp contrast with analysis of N-glycan that can be prepared by PNGase F, O-glycan analysis remains challenging due to a lack of versatile and simple procedures, especially those mediating cleavage of O-glycans from proteins. Most N-glycans and O-glycans are modified with sialic acids at the non-reducing end and their glycosidic linkages are labile, making it difficult to measure glycans by mass spectrometric analysis. In addition, sialic acid residues present on glycan chains via α2,3-, α2,6-, and α2,8-linkages as structural isomers.
Results: In this study, we firstly established a direct and linkage-specific derivatization method for sialylated O-glycans on proteins via linkage-specific lactone-opening aminolysis. In this procedure, labile sialylated glycans were not only stabilized, but also allowed distinguishing between sialyl linkages. Furthermore, we revealed that general reductive β-elimination was not useful for O-glycan cleavages with undesirable degradations of resulting methyl amides. Using β-elimination in the presence of pyrazolone (PMP), with low pH despite alkali base concentration, SALSA-derivatized O-glycans could be cleaved with minimal degradations. Cleaved and PMP-labeled O-glycans could be efficiently prepared in an open reaction system at high temperature (evaporative BEP reaction) and detected by simple liquid-phase extraction. Moreover, in the evaporative BEP reaction by changing the alkali solution with LiOH, the lithiated O-glycans could be observed and provided a lot of fragment information reflecting the complex structure of the O-glycans.
Significance: Direct sialic acid linkage-specific derivatization of O-glycans on glycoproteins is simple protocol containing in-solution aminolysis-SALSA and acetonitrile precipitation for removal of excess reagents. Evaporative β-elimination with pyrazolone makes possible intact O-linked glycan analysis just by liquid-phase extraction. These analytical methods established by the appropriate combination of direct-SALSA and evaporative β-elimination will facilitate O-glycomic studies in various biological samples.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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