Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays
Abstract Cellular glycosylation is characterized by chemical complexity and heterogeneity, which is challenging to reproduce synthetically. Here we show chemoenzymatic synthesis on phage to produce a genetically-encoded liquid glycan array (LiGA) of complex type N-glycans. Implementing the approach...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-08-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-40900-y |
| _version_ | 1797558190479507456 |
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| author | Chih-Lan Lin Mirat Sojitra Eric J. Carpenter Ellen S. Hayhoe Susmita Sarkar Elizabeth A. Volker Chao Wang Duong T. Bui Loretta Yang John S. Klassen Peng Wu Matthew S. Macauley Todd L. Lowary Ratmir Derda |
| author_facet | Chih-Lan Lin Mirat Sojitra Eric J. Carpenter Ellen S. Hayhoe Susmita Sarkar Elizabeth A. Volker Chao Wang Duong T. Bui Loretta Yang John S. Klassen Peng Wu Matthew S. Macauley Todd L. Lowary Ratmir Derda |
| author_sort | Chih-Lan Lin |
| collection | DOAJ |
| description | Abstract Cellular glycosylation is characterized by chemical complexity and heterogeneity, which is challenging to reproduce synthetically. Here we show chemoenzymatic synthesis on phage to produce a genetically-encoded liquid glycan array (LiGA) of complex type N-glycans. Implementing the approach involved by ligating an azide-containing sialylglycosyl-asparagine to phage functionalized with 50–1000 copies of dibenzocyclooctyne. The resulting intermediate can be trimmed by glycosidases and extended by glycosyltransferases yielding a phage library with different N-glycans. Post-reaction analysis by MALDI-TOF MS allows rigorous characterization of N-glycan structure and mean density, which are both encoded in the phage DNA. Use of this LiGA with fifteen glycan-binding proteins, including CD22 or DC-SIGN on cells, reveals optimal structure/density combinations for recognition. Injection of the LiGA into mice identifies glycoconjugates with structures and avidity necessary for enrichment in specific organs. This work provides a quantitative evaluation of the interaction of complex N-glycans with GBPs in vitro and in vivo. |
| first_indexed | 2024-03-10T17:26:53Z |
| format | Article |
| id | doaj.art-da21baab0c17465c9c943d4e2e9add04 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-10T17:26:53Z |
| publishDate | 2023-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-da21baab0c17465c9c943d4e2e9add042023-11-20T10:09:57ZengNature PortfolioNature Communications2041-17232023-08-0114111510.1038/s41467-023-40900-yChemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arraysChih-Lan Lin0Mirat Sojitra1Eric J. Carpenter2Ellen S. Hayhoe3Susmita Sarkar4Elizabeth A. Volker5Chao Wang6Duong T. Bui7Loretta Yang8John S. Klassen9Peng Wu10Matthew S. Macauley11Todd L. Lowary12Ratmir Derda13Department of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Molecular Medicine, The Scripps Research InstituteDepartment of Chemistry, University of AlbertaLectenz BioDepartment of Chemistry, University of AlbertaDepartment of Molecular Medicine, The Scripps Research InstituteDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaAbstract Cellular glycosylation is characterized by chemical complexity and heterogeneity, which is challenging to reproduce synthetically. Here we show chemoenzymatic synthesis on phage to produce a genetically-encoded liquid glycan array (LiGA) of complex type N-glycans. Implementing the approach involved by ligating an azide-containing sialylglycosyl-asparagine to phage functionalized with 50–1000 copies of dibenzocyclooctyne. The resulting intermediate can be trimmed by glycosidases and extended by glycosyltransferases yielding a phage library with different N-glycans. Post-reaction analysis by MALDI-TOF MS allows rigorous characterization of N-glycan structure and mean density, which are both encoded in the phage DNA. Use of this LiGA with fifteen glycan-binding proteins, including CD22 or DC-SIGN on cells, reveals optimal structure/density combinations for recognition. Injection of the LiGA into mice identifies glycoconjugates with structures and avidity necessary for enrichment in specific organs. This work provides a quantitative evaluation of the interaction of complex N-glycans with GBPs in vitro and in vivo.https://doi.org/10.1038/s41467-023-40900-y |
| spellingShingle | Chih-Lan Lin Mirat Sojitra Eric J. Carpenter Ellen S. Hayhoe Susmita Sarkar Elizabeth A. Volker Chao Wang Duong T. Bui Loretta Yang John S. Klassen Peng Wu Matthew S. Macauley Todd L. Lowary Ratmir Derda Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays Nature Communications |
| title | Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays |
| title_full | Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays |
| title_fullStr | Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays |
| title_full_unstemmed | Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays |
| title_short | Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays |
| title_sort | chemoenzymatic synthesis of genetically encoded multivalent liquid n glycan arrays |
| url | https://doi.org/10.1038/s41467-023-40900-y |
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