Biology subjects

Weatherill, E. E.

Publications and source records attributed to Weatherill, E. E..

2 recordsLinked to original sources

bioRxiv2021

Single-molecule imaging of cholesterol-dependent cytolysin assembly

We exploit single-molecule tracking and optical single channel recording in droplet interface bilayers to resolve the assembly pathway and pore-formation of the archetypical cholesterol-dependent cytolysin nanopore, Perfringolysin O. We follow the stoichiometry and diffusion of Perfringolysin O complexes during assembly with 60 millisecond temporal resolution and 20 nanometre spatial precision. Our results suggest individual nascent complexes can insert into the lipid membrane where they continue active assembly. Overall, these data support a model of stepwise irreversible assembly dominated by monomer addition, but with infrequent assembly from larger partial complexes.

biophysics↗

bioRxiv2021

Fast slow folding of an Outer Membrane Porin

In comparison to globular proteins, the spontaneous folding and insertion of {beta}-barrel membrane proteins is surprisingly slow, typically occurring on the order of minutes. Using single-molecule Forster Resonance Energy Transfer to report on the folding of fluorescently-labelled Outer Membrane Protein G we measured the real-time insertion of a {beta}-barrel membrane protein from an unfolded state. Folding events were rare, and fast (<20 ms); occurring immediately upon arrival at the membrane. This combination of infrequent, but rare, folding resolves this apparent dichotomy between slow ensemble kinetics, and the typical timescales of biomolecular folding.

biophysics↗