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Single-molecule imaging of glycan–lectin interactions on cells with Glyco-PAINT

Author(s)

• R. Riera, T.P. Hogervorst, W. Doelman, Y. Ni, S. Pujals, E. Bolli, J.D.C. Codée, S. I. van Kasteren & L. Albertazzi

Sources

Nature Chemical Biology (2021) https://doi.org/10.1038/s41589-021-00896-2

Most lectins bind carbohydrate ligands with relatively low affinity, making the identification of optimal ligands challenging. The authors 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. The diffusion coefficient of the receptor–sugar complex is also established.
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The uptake of ligands correlates with their binding affinity and residence time to establish the structure-function relations for various synthetic glycans. The authors 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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