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Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels
Metabolic glycoengineering (MGE) is an established method to incorporate chemical reporter groups into cellular glycans for subsequent bioorthogonal labeling. The method has found broad application for the visualization and isolation of glycans allowing their biological roles to be probed. Furthermo...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8048827/ https://www.ncbi.nlm.nih.gov/pubmed/33180370 http://dx.doi.org/10.1002/cbic.202000715 |
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author | Dold, Jeremias E. G. A. Wittmann, Valentin |
author_facet | Dold, Jeremias E. G. A. Wittmann, Valentin |
author_sort | Dold, Jeremias E. G. A. |
collection | PubMed |
description | Metabolic glycoengineering (MGE) is an established method to incorporate chemical reporter groups into cellular glycans for subsequent bioorthogonal labeling. The method has found broad application for the visualization and isolation of glycans allowing their biological roles to be probed. Furthermore, targeting of drugs to cancer cells that present high concentrations of sialic acids on their surface is an attractive approach. We report the application of a labeling reaction using 1,2‐diamino‐4,5‐methylenedioxybenzene for the quantification of sialic acid derivates after MGE with various azide‐ and alkene‐modified ManNAc, GlcNAc, and GalNAc derivatives. We followed the time course of sialic acid production and were able to detect sialic acids modified with the chemical reporter group – not only after addition of ManNAc derivatives to the cell culture. A cyclopropane‐modified ManNAc derivative, being a model for the corresponding cyclopropene analog, which undergoes fast inverse‐electron‐demand Diels‐Alder reactions with 1,2,4,5‐tetrazines, resulted in the highest incorporation efficiency. Furthermore, we investigated whether feeding the cells with natural and unnatural ManNAc derivative results in increased levels of sialic acids and found that this is strongly dependent on the investigated cell type and cell fraction. For HEK 293T cells, a strong increase in free sialic acids in the cell interior was found, whereas cell‐surface sialic acid levels are only moderately increased. |
format | Online Article Text |
id | pubmed-8048827 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-80488272021-04-20 Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels Dold, Jeremias E. G. A. Wittmann, Valentin Chembiochem Full Papers Metabolic glycoengineering (MGE) is an established method to incorporate chemical reporter groups into cellular glycans for subsequent bioorthogonal labeling. The method has found broad application for the visualization and isolation of glycans allowing their biological roles to be probed. Furthermore, targeting of drugs to cancer cells that present high concentrations of sialic acids on their surface is an attractive approach. We report the application of a labeling reaction using 1,2‐diamino‐4,5‐methylenedioxybenzene for the quantification of sialic acid derivates after MGE with various azide‐ and alkene‐modified ManNAc, GlcNAc, and GalNAc derivatives. We followed the time course of sialic acid production and were able to detect sialic acids modified with the chemical reporter group – not only after addition of ManNAc derivatives to the cell culture. A cyclopropane‐modified ManNAc derivative, being a model for the corresponding cyclopropene analog, which undergoes fast inverse‐electron‐demand Diels‐Alder reactions with 1,2,4,5‐tetrazines, resulted in the highest incorporation efficiency. Furthermore, we investigated whether feeding the cells with natural and unnatural ManNAc derivative results in increased levels of sialic acids and found that this is strongly dependent on the investigated cell type and cell fraction. For HEK 293T cells, a strong increase in free sialic acids in the cell interior was found, whereas cell‐surface sialic acid levels are only moderately increased. John Wiley and Sons Inc. 2020-12-17 2021-04-06 /pmc/articles/PMC8048827/ /pubmed/33180370 http://dx.doi.org/10.1002/cbic.202000715 Text en © 2020 The Authors. ChemBioChem published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Full Papers Dold, Jeremias E. G. A. Wittmann, Valentin Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels |
title | Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels |
title_full | Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels |
title_fullStr | Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels |
title_full_unstemmed | Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels |
title_short | Metabolic Glycoengineering with Azide‐ and Alkene‐Modified Hexosamines: Quantification of Sialic Acid Levels |
title_sort | metabolic glycoengineering with azide‐ and alkene‐modified hexosamines: quantification of sialic acid levels |
topic | Full Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8048827/ https://www.ncbi.nlm.nih.gov/pubmed/33180370 http://dx.doi.org/10.1002/cbic.202000715 |
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