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

Abstract

The objective of this critical review is to provide an overview of how emerging bioanalytical techniques are expanding our understanding of the complex physicochemical nature of virus interactions with host cell surfaces. Herein, selected model viruses representing both non-enveloped (simian virus 40 and human norovirus) and enveloped (influenza A virus, human herpes simplex virus, and human immunodeficiency virus type 1) viruses are highlighted. The technologies covered utilize a wide range of cell membrane mimics, from supported lipid bilayers (SLBs) containing a single purified host membrane component to SLBs derived from the plasma membrane of a target cell, which can be compared with live-cell experiments to better understand the role of individual interaction pairs in virus attachment and entry. These platforms are used to quantify binding strengths, residence times, diffusion characteristics, and binding kinetics down to the single virus particle and single receptor, and even to provide assessments of multivalent interactions. The technologies covered herein are surface plasmon resonance (SPR), quartz crystal microbalance with dissipation (QCM-D), dynamic force spectroscopy (DFS), total internal reflection fluorescence (TIRF) microscopy combined with equilibrium fluctuation analysis (EFA) and single particle tracking (SPT), and finally confocal microscopy using multi-labeling techniques to visualize entry of individual virus particles in live cells. Considering the growing scientific and societal needs for untangling, and interfering with, the complex mechanisms of virus binding and entry, we hope that this review will stimulate the community to implement these emerging tools and strategies in conjunction with more traditional methods. The gained knowledge will not only contribute to a better understanding of the virus biology, but may also facilitate the design of effective inhibitors to block virus entry.

Glycan Recognition in Human Norovirus Infections

“…Physiologically relevant culture systems such as HIEs and emerging bioanalytical techniques are enabling new understanding of virus-glycan interactions. As detailed in this review, this is very well defined in the case of HBGAs and HuNoVs and the body of evidence is ever-expanding for both HuNoVs and other pathogens [139]. The vast diversity of glycan recognition by different strains of HuNoV and among GII.4 variants is indicative of the elegance of viral evolution expanding the potential host repertoire.…”

Membrane attachment and fusion of HIV-1, influenza A, and SARS-CoV-2: resolving the mechanisms with biophysical methods

“…A range of analytical and biophysical techniques have been employed by virologists and biophysicists to resolve the molecular and cellular mechanism of the membrane attachment and fusion of the enveloped viruses under consideration (Harrison 2008 ; Hamilton et al 2012 ; Parveen et al 2018 ; Liu et al 2020 ; Bally et al 2021 ). Biochemical methods like enzyme-linked immunoassay and biophysical ensemble-averaged techniques like surface plasmon resonance (SPR) have been widely applied in characterizing the spike proteins and evaluating their binding affinity to membrane receptors (Myszka et al 2000 ; Suenaga et al 2012 ; Kim et al 2020 ; Walls et al 2020 ).…”

A cell free biomembrane platform for multimodal study of influenza virus hemagglutinin and for evaluation of entry-inhibitors against hemagglutinin

“…Importantly, multivalency pervades the mechanisms of viral-receptor interaction pathways, where the additive thermodynamic impact of multiple weak interactions promotes avidity and selectivity in receptor binding in host-virus interactions ( Mammenet al, 1998 ). This multivalency influences the motion of virus particles at the host cell surface ( Bally et al, 2021 ), as well as aiding in membrane reorganization. However, multivalent interactions make the overall viral-entry process so complex that it is extremely difficult to therapeutically inhibit the viral-cell entry.…”