%0 Figure %A Guo, Hongbo %A Rabouw, Huib %A Slomp, Anne %A Dai, Meiling %A van der Vegt, Floor %A van Lent, Jan W. M. %A McBride, Ryan %A Paulson, James C. %A J. de Groot, Raoul %A J. M. van Kuppeveld, Frank %A de Vries, Erik %A A. M. de Haan, Cornelis %D 2018 %T Kinetic model showing the basic flow of events taking place during virus particle binding. %U https://plos.figshare.com/articles/figure/Kinetic_model_showing_the_basic_flow_of_events_taking_place_during_virus_particle_binding_/6962339 %R 10.1371/journal.ppat.1007233.g008 %2 https://plos.figshare.com/ndownloader/files/12770390 %K epithelial host cells %K vivo IAV particles %K endpoint binding assays %K receptor-containing surfaces Interactions %K equilibrium binding models %K biolayer interferometric analysis %K mucus layer %K Multiple low-affinity HA-SIA interactions %K mucus decoy receptors %K Quantitative BLI analysis %K NA activity %K virus %K epithelial cell surfaces %X

Schematic representation of virus binding to a receptor-coated surface. Part of the viral envelope, containing HA (red symbol) and NA (blue symbol), and the sensor surface, coated with SIA (purple diamond)-containing receptors (R), is shown. Kinetically different steps lead to multivalent interaction between IAV and a receptor-coated surface. The initial HA-dependent binding event (step 1 in red) is a virus concentration-dependent intermolecular process governed by a binding rate constant kon (with the unit M−1 s−1) and a dissociation constant koff (with the unit s−1). Subsequent HA binding steps (e.g. steps 2 and 4) are intramolecular with a kon (not necessarily the same as in the first step) and koff (both with the unit s−1). For IAV, having a KD (koff/kon) of ~0.3 to ~3mM for a monovalent HA-receptor interaction, binding at pM concentrations inevitably results in a low binding rate that is mostly determined by the first binding event. NA can also contribute to the initial binding rate. This contributory effect can be inhibited by OC and is therefore attributed to binding of receptor to a NA catalytic site. The contribution of NA to receptor binding is determined by a dissociation constant (KD = koff/koff) for the substrate (step I in blue) and a catalytic rate constant (kcat; bold blue arrow) determining the receptor cleavage rate. A lower kcat will result in prolonged receptor binding before cleavage or dissociation takes place. This will enhance the chance for additional binding events (mostly by HA, which is present at higher density than NA), thereby promoting the cascade of multivalent interactions responsible for tight virus binding. Given the lower KD (30μM~600μM) [51] of NA, in comparison to HA, for interaction with sialosides, a considerable contribution to the initial binding rate by NA is expected even whereas the NA/HA ratio of a virus particle is generally quite low. Longer lasting, BLI-detectable, binding requires the formation of additional HA- and/or NA–SIA bonds, which is indicated by the grey shaded area. Initial binding events will be hardly detected due to the low levels of equilibrium binding in step 1 and I. During the BLI-detectable phase of binding, HA- and NA–SIA interactions are formed and broken in a virus concentration independent mode with the result that all binding states can rapidly interconvert via binding/dissociation events 2 to 11 and II to IV. The number of simultaneous interactions that can occur is logically dependent on receptor density and koff/kon ratios but how many simultaneous interactions suffice to keep a virus particle bound to the surface remains unknown. The experiments shown in Fig 1 suggest the number of interactions required is very low. Theoretically, the dynamics of HA-SIA interactions allow a virus to roll over the surface but experiments shown in Fig 7 show that NA activity strongly stimulates rolling (and eventually leads to virus dissociation). This is schematically indicated by the curled arrows where NA cleavage activity creates receptor-free positions on the surface. The receptor gradient caused in this way is probably the driving force for virus rolling but the direction in which the virus rolls (away from the empty position or “reaching over” the empty position) still needs further research.

%I PLOS Pathogens