Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures

Abstract Glycans in tissues are structurally diverse and usually include a large number of isomers that cannot be easily distinguished by mass spectrometry (MS). To address this issue, we developed a combined method that can efficiently separate and identify glycan isomers. First, we separated 2-ami...

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Autores principales:	Noriko Suzuki, Tatsuya Abe, Ken Hanzawa, Shunji Natsuka
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8f3aad031a6042d7841c7e943f01c5ae
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spelling	oai:doaj.org-article:8f3aad031a6042d7841c7e943f01c5ae2021-12-02T16:31:13ZToward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures10.1038/s41598-021-84668-x2045-2322https://doaj.org/article/8f3aad031a6042d7841c7e943f01c5ae2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-84668-xhttps://doaj.org/toc/2045-2322Abstract Glycans in tissues are structurally diverse and usually include a large number of isomers that cannot be easily distinguished by mass spectrometry (MS). To address this issue, we developed a combined method that can efficiently separate and identify glycan isomers. First, we separated 2-aminopyridine (PA)-derivatized N-glycans from chicken colon by reversed-phase liquid chromatography (LC) and directly analyzed them by electrospray ionization (ESI)-MS and MS/MS to obtain an overview of the structural features of tissue glycans. Next, we deduced the structures of isomers based on their elution positions, full MS, and MS/MS data, before or after digestions with several exoglycosidases. In this method, the elution position differed greatly depending on the core structure and branching pattern, allowing multiantennary N-glycan structures to be easily distinguished. To further determine linkages of branch sequences, we modified PA-N-glycans with sialic acid linkage-specific alkylamidation and/or permethylation, and analyzed the products by LC–MS and multistage MS. We determined the relative abundances of core structures, branching patterns, and branch sequences of N-glycans from chicken colon, and confirmed presence of characteristic branch sequences such as Lex, sialyl Lex, sulfated LacNAc, LacNAc repeat, and LacdiNAc. The results demonstrated that our method is useful for comparing N-glycomes among various tissue samples.Noriko SuzukiTatsuya AbeKen HanzawaShunji NatsukaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Noriko Suzuki Tatsuya Abe Ken Hanzawa Shunji Natsuka Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures
description	Abstract Glycans in tissues are structurally diverse and usually include a large number of isomers that cannot be easily distinguished by mass spectrometry (MS). To address this issue, we developed a combined method that can efficiently separate and identify glycan isomers. First, we separated 2-aminopyridine (PA)-derivatized N-glycans from chicken colon by reversed-phase liquid chromatography (LC) and directly analyzed them by electrospray ionization (ESI)-MS and MS/MS to obtain an overview of the structural features of tissue glycans. Next, we deduced the structures of isomers based on their elution positions, full MS, and MS/MS data, before or after digestions with several exoglycosidases. In this method, the elution position differed greatly depending on the core structure and branching pattern, allowing multiantennary N-glycan structures to be easily distinguished. To further determine linkages of branch sequences, we modified PA-N-glycans with sialic acid linkage-specific alkylamidation and/or permethylation, and analyzed the products by LC–MS and multistage MS. We determined the relative abundances of core structures, branching patterns, and branch sequences of N-glycans from chicken colon, and confirmed presence of characteristic branch sequences such as Lex, sialyl Lex, sulfated LacNAc, LacNAc repeat, and LacdiNAc. The results demonstrated that our method is useful for comparing N-glycomes among various tissue samples.
format	article
author	Noriko Suzuki Tatsuya Abe Ken Hanzawa Shunji Natsuka
author_facet	Noriko Suzuki Tatsuya Abe Ken Hanzawa Shunji Natsuka
author_sort	Noriko Suzuki
title	Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures
title_short	Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures
title_full	Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures
title_fullStr	Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures
title_full_unstemmed	Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures
title_sort	toward robust n-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8f3aad031a6042d7841c7e943f01c5ae
work_keys_str_mv	AT norikosuzuki towardrobustnglycomicsofvarioustissuesamplesthatmaycontainglycanswithunknownorunexpectedstructures AT tatsuyaabe towardrobustnglycomicsofvarioustissuesamplesthatmaycontainglycanswithunknownorunexpectedstructures AT kenhanzawa towardrobustnglycomicsofvarioustissuesamplesthatmaycontainglycanswithunknownorunexpectedstructures AT shunjinatsuka towardrobustnglycomicsofvarioustissuesamplesthatmaycontainglycanswithunknownorunexpectedstructures
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Toward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structures

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