Capturing fusion glycoproteins in motion: a focus on Ebola virus

The Ebola virus (EBOV) fusion glycoprotein GP mediates cell entry and is the main virus antigenic target. The molecular mechanisms underlying entry and antibody-mediated neutralization remain largely unknown, but are crucial for developing efficient therapies. To uncover these mechanisms, I will develop cutting-edge biophysical approaches that capture GP structural dynamics and host interactions during the EBOV lifecycle on membrane surfaces reflecting the authentic virus, with the overall aim of designing alternative candidate antivirals. I will establish a hydrogen-deuterium exchange mass spectrometry (HDX-MS) platform capable of tracking the conformational trajectory and molecular switches of GP embedded in virus-like particles undergoing the step- wise transitional events leading to EBOV entry (pH changes, lysosomal proteolysis and receptor binding). I will combine dynamic mass photometry with HDX-MS to derive a full picture of how human monoclonal antibodies engage GP, at the conformational and macro-molecular level, deciphering antibody multivalency, binding mechanisms and the immune response synergy that underlie EBOV neutralization. I will exploit these insights to develop data-driven computational approaches to rationally design GP-targeting neutralizing nanobodies of potential therapeutic success. This integrated approach will be both unique and broadly applicable to study the infectious processes of emerging and established enveloped viruses, thus contributing to pandemic preparedness.