Multiple recent reports indicate that the S protein of SARS-CoV-2 specifically interacts with membrane receptors and attachment factors other than ACE2. They likely have an active role in cellular attachment and entry of the virus. In this article, we examined the binding of SARS-CoV-2 particles to gangliosides embedded in supported lipid bilayers (SLBs), mimicking the cell membrane-like environment. We show that the virus specifically binds to sialylated (sialic acid (SIA)) gangliosides, i.e., GD1a, GM3, and GM1, as determined from the acquired single-particle fluorescence images using a time-lapse total internal reflection fluorescence (TIRF) microscope. The data of virus binding events, the apparent binding rate constant, and the maximum virus coverage on the ganglioside-rich SLBs show that the virus particles have a higher binding affinity toward the GD1a and GM3 compared to the GM1 ganglioside. Enzymatic hydrolysis of the SIA−Gal bond of the gangliosides confirms that the SIA sugar unit of GD1a and GM3 is essential for virus attachment to the SLBs and even the cell surface sialic acid is critical for the cellular attachment of the virus. The structural difference between GM3/GD1a and GM1 is the presence of SIA at the main or branched chain. We conclude that the number of SIA per ganglioside can weakly influence the initial binding rate of SARS-CoV-2 particles, whereas the terminal or more exposed SIA is critical for the virus binding to the gangliosides in SLBs.

The present work aims to study the fluidic behavior on lateral confinement by placing sidewalls on the planar plug nozzle through experiments. This study involves two cases of nozzle pressure ratio (NPR = 3, 6), which correspond to over-expanded nozzle operating conditions. Steady-state pressure measurements, together with schlieren and surface oil flow visualization, reveal the presence of over-expansion shock and subsequent interaction and modification of the flow field on the plug surface. The flow remains attached to the plug surface for NPR = 3; whereas for NPR = 6, a separated flow field with a recirculation bubble is observed. Spectral analysis of the unsteady pressure signals illustrates a clear difference between the attached and the separated flow. Besides, other flow features with a distinct temporal mode associated with and without lateral confinement are observed. The absence of lateral confinement reduces the intensity of low-frequency unsteadiness; however, on the contrary, the interaction region is relatively reduced under lateral confinement.

Gangliosides, forming a class of lipids complemented by sugar chains, influence the lateral distribution of membrane proteins or membrane-binding proteins, act as receptors for viruses and bacterial toxins, and mediate several types of cellular signaling. Gangliosides incorporated into supported lipid bilayers (SLBs) have been widely applied as a model system to examine these biological processes. In this work, we explored how ganglioside composition affects the kinetics of SLB formation using the vesicle rupturing method on a solid surface. We imaged the attachment of vesicles and the subsequent SLB formation using the time-lapse total internal reflection fluorescence microscopy technique. In the early phase, the ganglioside type and concentration influence the adsorption kinetics of vesicles and their residence/lifetime on the surface before rupturing. Our data confirm that a simultaneous rupturing of neighboring surface-adsorbed vesicles forms microscopic lipid patches on the surface and it is triggered by a critical coverage of the vesicles independent of their composition. In the SLB growth phase, lipid patches merge, forming a continuous SLB. The propagation of patch edges catalyzes the process and depends on the ganglioside type. Our pH-dependent experiments confirm that the polar/charged head groups of the gangliosides have a critical role in these steps and phases of SLB formation kinetics.

The present work aims to study the fluidic behavior on lateral confinement by placing side-walls on the planar plug nozzle through experiments. The study involves two cases of nozzle pressure ratio (NPR=3, 6), which correspond to over-expanded nozzle operating conditions. Steady-state pressure measurements, together with schlieren and surface oil flow visualization, reveal the presence of over-expansion shock and subsequent interaction and modification of the flow field on the plug surface. The flow remains attached to the plug surface for NPR=3; whereas, for NPR=6, a separated flow field with a recirculation bubble is observed. Spectral analysis of the unsteady pressure signals illustrates a clear difference between the attached and the separated flow. Besides, other flow features with a distinct temporal mode associated with and without lateral confinement are observed. The absence of lateral confinement reduces the intensity of low-frequency unsteadiness; however, on the contrary, the interaction region is relatively reduced under lateral confinement.