Guo, Y, Nehlmeier, I, Poole, E et al. (12 more authors) (2017) Dissecting Multivalent Lectin-Carbohydrate Recognition Using Polyvalent Multifunctional Glycan-Quantum Dots. Journal of the American Chemical Society, 139 (34). pp. 11833-11844. ISSN 0002-7863
Abstract
Multivalent protein-carbohydrate interactions initiate the first contacts between virus/bacteria and target cells which ultimately lead to infection. Understanding the structures and binding modes involved is vital to the design of specific, potent multivalent inhibitors. However, the lack of structural information on such flexible, complex and multimeric cell surface membrane proteins has often hampered such endeavours. Herein we report that quantum dots (QDs) displayed with a dense array of mono-/di-saccharides are powerful probes for multivalent protein-glycan interactions. Using a pair of closely related tetrameric lectins, DC-SIGN and DC-SIGNR, which bind to the HIV and Ebola virus glycoproteins (EBOV-GP) to augment viral entry and infect target cells, we show that such QDs efficiently dissect the different DC-SIGN/R-glycan binding modes (tetra-/di-/mono- valent) through a combination of multi-modal readouts: Förster resonance energy transfer (FRET), hydrodynamic size measurement and transmission electron microscopy imaging. We also report a new QD-FRET method for quantifying QD-DC-SIGN/R binding affinity, revealing that DC-SIGN binds to the QD >100 fold tighter than DC-SIGNR. This result is consistent with DC-SIGN’s higher trans-infection efficiency of some HIV strains over DC-SIGNR. Finally, we show that the QDs potently inhibit DC-SIGN-mediated enhancement of EBOV-GP-driven transduction of target cells with IC50 values down to 0.7 nM, matching well to their DC-SIGN binding constant (apparent Kd = 0.6 nM) measured by FRET. These results suggest that the glycan-QDs are powerful multifunctional probes for dissecting multivalent protein-ligand recognition and predicting glyconanoparticle inhibition of virus infection at the cellular level.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | © 2017 American Chemical Society. This document is the Accepted Manuscript version of a Published Work that appeared in final form in Journal of the American Chemical Society, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/jacs.7b05104. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Quantum dot; DC-SIGN; Multivalency; multi-modal readout; glyconanoparticle; virus inhibition; Förster resonance energy transfer |
Dates: |
|
Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemical & Process Engineering (Leeds) The University of Leeds > Faculty of Environment (Leeds) > School of Food Science and Nutrition (Leeds) The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) > Inorganic Chemistry (Leeds) The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) > Organic Chemistry (Leeds) |
Funding Information: | Funder Grant number BBSRC BB/M005666/1 Wellcome Trust 097354/Z/11/Z EPSRC EP/M028143/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 10 Aug 2017 11:52 |
Last Modified: | 08 Aug 2018 00:38 |
Status: | Published |
Publisher: | American Chemical Society |
Identification Number: | 10.1021/jacs.7b05104 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:119958 |