A Structural Study on the Rabies Virus Glycoprotein combining Cryo-EM and Mass Spectrometry-based Approaches
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Rabies Virus (RABV) is a zoonotic neurotropic virus and the main causative agent for Rabies, a lethal encephalitis. The Rabies Virus Glycoprotein (RABV G) is a homo-trimer present on the viral envelope and is essential in the viral life cycle because of its role in receptor recognition and membrane fusion and is the main immunogenic target. Here we report an updated protocol for the purification of RABV G from cell supernatant, which was used for cryo-EM single particle analysis to elucidate its structure. Additionally we mapped the N-linked glycans of the protein and characterized the oligomerization using mass photometry, native MS and Charge-Detection MS, which hinted at a concentration-dependent equilibrium. We also assessed the binding of two therapeutically used neutralizing antibodies by HDX-MS and can point to a probable epitope on RABV G for both.