Fusion between host cell membranes and viral envelopes remains one of the most crucial steps in viral pathogenesis and allows enveloped viruses to overcome one of the largest barriers to propagation: cell entry. Various aspects of fusion mechanics have been analysed using techniques such as electron microscopy (EM), cryo-electron tomography (cryo-ET), and sedimentation equilibrium analytical ultracentrifugation. However, we propose the novel use of liposome-coated iron oxide nanoparticles (LIONs) as a new approach for the analysis of viral fusion mechanisms. We demonstrate the feasibility of using LIONs for the analysis of fusion using the well-understood fusion mechanics of influenza. Hemagglutinin (HA), a fusion glycoprotein is known to play an important role in this process. LIONs are synthesized by coating iron oxide nanoparticles and fluorescent dye in a liposomal membrane, which effectively mimics a host endosomal membrane. This platform, and the dual magnetic and fluorescent signals it provides, may then be used to observe interactions between the membrane coating and fusion proteins in real-time. In this presentation, we will demonstrate the ability of LIONs to observe fusion in real-time between LION membranes and HA/ HA2 fusion peptides as well as influenza virus-like particles (VLPs). In addition to this, we also use the LION platform to study environmental effects on fusion, including pH, membrane cholesterol composition, presence of the protease trypsin and small molecule inhibitor arbidol.<br/>

Liposome-coated iron oxide nanoparticles (LIONs): A dynamic approach for the investigation of influenza fusion mechanisms

2019 Spring National Meeting

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