Competition for Membrane Receptors: Norovirus Detachment via Lectin Attachment

Abstract

Virus internalization into the host cells occurs via multivalent interactions, in which a single virus binds to multiple receptors in parallel. Because of analytical and experimental limitations this complex type of interaction is still poorly understood and quantified. Herein, the multivalent interaction of norovirus-like particles (noroVLPs) with H or B type 1 glycosphingolipids (GSLs), embedded in a supported phospholipid bilayer, is investigated by following the competition between noroVLPs and a lectin (from Ralstonia solanacearum) upon binding to these GSLs. Changes in noroVLP and lectin coverage, caused by competition, were monitored for both GSLs and at different GSL concentrations using quartz crystal microbalance with dissipation monitoring. The study yields information about the minimum GSL concentration needed for i) noroVLPs to achieve firm attachment to the bilayer prior to competition, and to ii) remain firmly attached to the bilayer during competition. We show that these two concentrations are almost identical for the H type 1-noroVLP interaction, but differ for B type 1, indicating an accumulation of B type 1 GSLs in the noroVLP-bilayer interaction area. Furthermore, the GSL concentration required for firm attachment is significantly larger for H type 1 than for B type 1, indicating a higher affinity of noroVLP towards B type 1. This finding is supported by extracting the energy of single noroVLP-H type 1 and noroVLP-B type 1 bonds from the competition kinetics, which were estimated to be 5 and 6 kcal/mol, respectively. This demonstrates the potential of utilizing competitive binding kinetics to analyse multivalent interactions, which has remained difficult to quantify using conventional approaches.

Complete assignment of Ala, Ile, LeuProS, Met and ValProS methyl groups of the protruding domain from human norovirus GII.4 Saga

“…CSPs were observed as far as 25 Å from the HBGA binding pocket at ligand saturation, suggesting the presence of an allosteric network regulated by HBGA binding. A recent study monitored the binding of human norovirus virus-like particles to glycosphingolipids embedded in a phospholipid bilayer (Parveen et al 2019). From competitive titrations, it was concluded that the intrinsic (per binding site) bond energies of H type 1 and B type 1 glycosphingolipids are in good agreement with the K D s reported for HBGAs by NMR.…”

Physicochemical tools for studying virus interactions with targeted cell membranes in a molecular and spatiotemporally resolved context

“…By solving the rate equations of CTxB binding and SV40 release and by using estimates for the maximum valency obtainable by SV40 on a planar membrane, it was possible to relate the observed release kinetics with the binding strength of the single VP1-GM1 pair. In a follow-up study, Parveen et al [ 78 ] applied the same methodology to probe binding of human norovirus VLPs to histo-blood group antigens (HBGAs) presented on GSLs and release of these VLPs from the HBGAs upon addition of a lectin from Ralstonia solanacearum (Fig. 3A, B ).…”

“…As the friction strongly depends on the size of the adsorbed object, it is possible to distinguish binding events caused by viruses and proteins and thus to probe the release of membrane-bound viruses upon addition of a binding competitor having a higher affinity to the receptor/attachment factor (e.g., a lectin). This provides information on the binding strength of the virus-membrane interaction and (B) on the accumulation of receptors/attachment factors by the virus, which generates deviations between the relative virus surface coverage, determined during the initial binding process (B, solid line) or lectin-induced virus release (B, circles) [ 78 ]. (C) TIRF microscopy can be used to probe the binding of receptor/attachment factor-containing, dye-labeled vesicles and interface-linked viruses, allowing to determine the impact of the constituents (e.g., vesicle composition or virus strain) on the overall interaction.…”

“…Thanks to such an experiment, the attachment rates (slopes in D, left) as well as the off-rates (decay of residence time, D right) of virus-membrane interactions can be related to the patterns of susceptibility to infection for various individuals [ 23 , 55 ]. Adapted figures with permission from [ 23 , 55 , 78 ]; Copyright by the American Chemical Society and American Physical Society, respectively …”

Determination of Nano-sized Adsorbate Mass in Solution using Mechanical Resonators: Elimination of the so far Inseparable Liquid Contribution

“…8 Still, the rich information contained in combined ∆f n and ∆D n measurements, especially when combined with theoretical models representing the response for viscoelastic films, 9,10 has turned out very valuable in multiple research areas, including hydration analysis of organic polymers 11,12 , proteins 13 and biological membranes 13,14 . The method is also widely applied to investigate material porosity 15,16 , growth of mesoporous material 17 , characterization of biomimetic membranes 18,19 , development of bioanalytical sensors [20][21][22][23] , as well as in studies of biomolecular interactions 24,25 , including interrogations of their structural 26 and orientational 27 changes.…”