Quantification of Multivalency in Protein-Oligomer-Coated Nanoparticles Targeting Dynamic Membrane Glycan Receptors.

Abstract

Multivalent binding of proteins to glycan receptors on the host cell quantitatively controls the initial adhesion of most viruses. However, quantifying such multivalency in terms of binding valency has always been a challenge because of the hierarchy of multivalency involving multiple protein oligomers on the virus, limiting our understanding of virus adhesion and virulence. To address this challenge, we mimicked virus adhesion to cell surfaces by attaching protein-oligomer-coated nanoparticles (NPs) to fluidic glycolipid membranes with surface glycan density varying over 4 orders of magnitude. Using total internal reflection fluorescence microscopy to track single attached NPs, we show that the binding isotherms exhibit two regions, attributed to monovalent and multivalent protein/glycan interactions at low and high glycan densities, respectively. The bimodal binding curve allows the quantification of the different valency and binding constants of monovalent and multivalent interactions. In addition, the competitive inhibition of multivalency by the glycopolymer presenting multiple glycan moieties is quantitatively appreciated. This work is essential to mapping and understanding the complex binding specificities of glycan-binding proteins and inhibitory drug designs and applications.