The C-type lectin receptor DC-SIGN has been highlighted as the coreceptor for the spike protein of the SARS-CoV-2 virus. A multivalent glycomimetic ligand, Polyman26, has been found to inhibit DC-SIGN-dependent trans-infection of SARS-CoV-2. The molecular details underlying avidity generation in such systems remain poorly characterized. In an effort to dissect the contribution of the known multivalent effects - chelation, clustering, and statistical rebinding - we studied a series of dendrimer constructs related to Polyman26 with a rod core rationally designed to engage simultaneously two binding sites of the tetrameric DC-SIGN. Binding properties of these compounds have been studied with a range of biophysical techniques, including recently developed surface plasmon resonance oriented-surface methodology. Using molecular modeling we addressed, for the first time, the impact of the carbohydrate recognition domains' flexibility of the DC-SIGN tetramer on the compounds' avidity. We were able to gain deeper insight into the role of different binding modes, which in combination produce a construct with a nanomolar affinity despite a limited valency. This multifaceted experimental-theoretical approach provides detailed understanding of multivalent ligand/multimeric protein interactions which can lead to future predictions. This work opens the way to the development of new virus attachment blockers adapted to different C-type lectin receptors of viruses.

Astbury Centre and School of Molecular and Cellular Biology University of Leeds Leeds LS2 9JT United Kingdom

Department of Physics and Astronomy Augusto Righi University of Bologna Via Zamboni 33 40126 Bologna Italy

Institut Universitaire de France 1 rue Descartes 75231 Paris France

Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences Flemingovo nam 2 Prague 6 166 10 Czechia

Univ Grenoble Alpes CNRS CEA Institut de Biologie Structurale 38000 Grenoble France

Univ Grenoble Alpes CNRS CERMAV 38000 Grenoble France

Universita' degli Studi di Milano Dipartimento di Chimica via Golgi 19 20133 Milano Italy

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