Single-molecule imaging of glycan-lectin interactions on cells with Glyco-PAINT
© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc..
Most lectins bind carbohydrate ligands with relatively low affinity, making the identification of optimal ligands challenging. Here we introduce a point accumulation in nanoscale topography (PAINT) super-resolution microscopy method to capture weak glycan-lectin interactions at the single-molecule level in living cells (Glyco-PAINT). Glyco-PAINT exploits weak and reversible sugar binding to directly achieve single-molecule detection and quantification in cells and is used to establish the relative kon and koff rates of a synthesized library of carbohydrate-based probes, as well as the diffusion coefficient of the receptor-sugar complex. Uptake of ligands correlates with their binding affinity and residence time to establish structure-function relations for various synthetic glycans. We reveal how sugar multivalency and presentation geometry can be optimized for binding and internalization. Overall, Glyco-PAINT represents a powerful approach to study weak glycan-lectin interactions on the surface of living cells, one that can be potentially extended to a variety of lectin-sugar interactions.
Medienart: |
E-Artikel |
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Erscheinungsjahr: |
2021 |
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Erschienen: |
2021 |
Enthalten in: |
Zur Gesamtaufnahme - volume:17 |
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Enthalten in: |
Nature chemical biology - 17(2021), 12 vom: 11. Dez., Seite 1281-1288 |
Sprache: |
Englisch |
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Beteiligte Personen: |
Riera, Roger [VerfasserIn] |
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Links: |
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Themen: |
Journal Article |
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Anmerkungen: |
Date Completed 28.12.2021 Date Revised 28.10.2022 published: Print-Electronic Citation Status MEDLINE |
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doi: |
10.1038/s41589-021-00896-2 |
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funding: |
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Förderinstitution / Projekttitel: |
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PPN (Katalog-ID): |
NLM333023641 |
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520 | |a Most lectins bind carbohydrate ligands with relatively low affinity, making the identification of optimal ligands challenging. Here we introduce a point accumulation in nanoscale topography (PAINT) super-resolution microscopy method to capture weak glycan-lectin interactions at the single-molecule level in living cells (Glyco-PAINT). Glyco-PAINT exploits weak and reversible sugar binding to directly achieve single-molecule detection and quantification in cells and is used to establish the relative kon and koff rates of a synthesized library of carbohydrate-based probes, as well as the diffusion coefficient of the receptor-sugar complex. Uptake of ligands correlates with their binding affinity and residence time to establish structure-function relations for various synthetic glycans. We reveal how sugar multivalency and presentation geometry can be optimized for binding and internalization. Overall, Glyco-PAINT represents a powerful approach to study weak glycan-lectin interactions on the surface of living cells, one that can be potentially extended to a variety of lectin-sugar interactions | ||
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700 | 1 | |a Doelman, Ward |e verfasserin |4 aut | |
700 | 1 | |a Ni, Yan |e verfasserin |4 aut | |
700 | 1 | |a Pujals, Silvia |e verfasserin |4 aut | |
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700 | 1 | |a Codée, Jeroen D C |e verfasserin |4 aut | |
700 | 1 | |a van Kasteren, Sander I |e verfasserin |4 aut | |
700 | 1 | |a Albertazzi, Lorenzo |e verfasserin |4 aut | |
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