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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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-84668-x |
| _version_ | 1818842922646241280 |
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| author | Noriko Suzuki Tatsuya Abe Ken Hanzawa Shunji Natsuka |
| author_facet | Noriko Suzuki Tatsuya Abe Ken Hanzawa Shunji Natsuka |
| author_sort | Noriko Suzuki |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-19T04:49:40Z |
| format | Article |
| id | doaj.art-8f3aad031a6042d7841c7e943f01c5ae |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-19T04:49:40Z |
| publishDate | 2021-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-8f3aad031a6042d7841c7e943f01c5ae2022-12-21T20:35:24ZengNature PortfolioScientific Reports2045-23222021-03-0111111510.1038/s41598-021-84668-xToward robust N-glycomics of various tissue samples that may contain glycans with unknown or unexpected structuresNoriko Suzuki0Tatsuya Abe1Ken Hanzawa2Shunji Natsuka3Graduate School of Science and Technology, Niigata UniversityGraduate School of Science and Technology, Niigata UniversityGraduate School of Science and Technology, Niigata UniversityGraduate School of Science and Technology, Niigata UniversityAbstract 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.https://doi.org/10.1038/s41598-021-84668-x |
| spellingShingle | 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 Scientific Reports |
| title | 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_short | 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 |
| url | https://doi.org/10.1038/s41598-021-84668-x |
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