The role
We have discovered a free fatty acid-binding pocket in SARS-CoV-2 spike which binds specifically LA, with nanomolar affinity (doi: 10.1126/science.abd3255). We showed that LA-binding induces a locked spike conformation incompatible with binding to human host cell receptor ACE2, thus inhibiting viral infection. LA treatment of human cells already infected with SARS-CoV-2 suppresses viral replication and results in deformed virions (doi: 10.1126/sciadv.adc9179). In the current project, we aim to elucidate the functional importance of the pocket and how it regulates viral replication. To obtain these fundamental new insights, we will use biochemical, biophysical and structural (cryo-EM) approaches to design and characterize SARS-CoV-2 spike protein mutants.
We will dissect the impact of LA-binding to the pocket on spike architecture and ACE2 receptor binding, and we will elucidate the effect of LA-treatment on viral infection and replication in cells with mutant virus (collaboration with Prof Andrew Davidson, Bristol).
We will analyse mutant virion morphology and spike conformation in situ(collaboration with Prof Paul Verkade, Bristol), using state-of-the-art imaging approaches and infrastructure available in Bristol (cryo-FIB-SEM, Talos Arctica microscope) in the GW4 Facility for High-Resolution Cryo-EMand at the national facility eBIC in Harwell via GW4 BAG access.
What will you be doing?
You will be functionally dissecting the free fatty acid-binding pocket in the SARS-CoV-2 Spike protein. To this end, you will produce SARS-CoV-2 spike mutants we designed (collaboration with Prof Adrian Mulholland, Bristol) using MultiBac insect cell expression, confirm that LA binding is abrogated, and characterise ACE2 receptor binding and stability of the spike mutant using biophysics. You will solve the structure of the best SARS-CoV-2 spike mutant protein by single particle cryo-EM and in situ in the context of mutant SARS-CoV-2 using cryo-tomography.
You should apply if
You hold, or expect to hold shortly, a PhD in Structural Biology, Biochemistry, Biophysics or related. You have published experience in protein expression and purification, biophysical characterization, Cryo-EM and image processing.
