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

Roger Riera, Tim P. Hogervorst, Ward Doelman, Yan Ni, Silvia Pujals, Evangelia Bolli, Jeroen D.C. Codée, Sander I. van Kasteren, Lorenzo Albertazzi*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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. [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)1281-1288
Number of pages8
JournalNature Chemical Biology
Volume17
Issue number12
DOIs
Publication statusPublished - Dec 2021
Externally publishedYes

Bibliographical note

Funding Information:
R.R. and L.A. thank the European Research Council/Horizon 2020 for financial support (no. ERC-StG-757397). L.A. thanks NWO for support (VIDI grant no. 192.028). L.A. thanks the Barcelona Institute of Science and Technology for support. This work was funded by the NWO gravitation program 2013 granted to the Institute for Chemical

Funding Information:
(4) and the European Research Council (grant no.ERC-CoG-865175,S.I.v.K.).Immunology (no.ICI-024.002.009)(T.P.H.andJ.D.C.C.);NWOBBoL grant(W.D.);

Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer Nature America, Inc.

Funding

R.R. and L.A. thank the European Research Council/Horizon 2020 for financial support (no. ERC-StG-757397). L.A. thanks NWO for support (VIDI grant no. 192.028). L.A. thanks the Barcelona Institute of Science and Technology for support. This work was funded by the NWO gravitation program 2013 granted to the Institute for Chemical (4) and the European Research Council (grant no.ERC-CoG-865175,S.I.v.K.).Immunology (no.ICI-024.002.009)(T.P.H.andJ.D.C.C.);NWOBBoL grant(W.D.);

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