How curvature-generating proteins build scaffolds on membrane nanotubes

Bin/Amphiphysin/Rvs (BAR) domain proteins control the curvature of lipid membranes in endocytosis, trafficking, cell motility, the formation of complex sub-cellular structures, and many other cellular phenomena. They form three-dimensional assemblies, which act as molecular scaffolds to reshape the membrane and alter its mechanical properties. It is unknown, however, how a protein scaffold forms and how BAR domains interact in these assemblies at protein densities relevant for a cell. In this work, we employ various experimental, theoretical and simulation approaches to explore how BAR proteins organize to form a scaffold on a membrane nanotube. By combining quantitative microscopy with analytical modeling, we demonstrate that a highly curving BAR protein endophilin nucleates its scaffolds at the ends of a membrane tube, contrary to a weaker curving protein centaurin, which binds evenly along the tube’s length. Our work implies that the nature of local protein-membrane interactions can affect the specific localization of proteins on membrane-remodeling sites. Furthermore, we show that amphipathic helices are dispensable in forming protein scaffolds. Finally, we explore a possible molecular structure of a BAR-domain scaffold using coarse-grained molecular dynamics simulations. Together with fluorescence microscopy, the simulations show that proteins need only to cover 30–40% of a tube’s surface to form a rigid assembly. Our work provides mechanical and structural insights into the way BAR proteins may sculpt the membrane as a high-order cooperative assembly in important biological processes. 

Aged embankment imaging and assessment using surface waves

Rapid, non-intrusive surface wave surveys provide depth profiles from which ground models can be generated for use in earthwork condition assessment. Stiffness throughout earthworks controls the behaviour under static and dynamic loads, and characterising heterogeneity is of interest in relation to the stability of engineered backfill and life-cycle deterioration in aged utility and transportation infrastructure. Continuous surface wave methods were used to identify interfaces between fine- and coarse-grained fill in an end-tipped embankment along the Great Central Railway in Nottinghamshire, UK. Multichannel analysis of surface wave (MASW) methods were used to characterise subsurface voiding in a canal embankment along the Knottingley and Goole canal near Eggborough, Yorkshire. MASW methods are currently being used to study extreme weather impacts on the stability of a highplasticity clay embankment along the Gloucestershire–Warwickshire railway near Laverton. Optimal results were obtained using equipment capable of generating and detecting over wide frequency ranges.