An in vitro gram-negative double bilayer envelope mimetic

In gram-negative bacteria the envelope is a complex, essential, tripartite structure formed by two membranes sandwiching a peptidoglycan layer that together maintain the integrity and shape of the cell. While the structure and composition of the bacterial cell envelope has been known for a number of years, emulating this system accurately in vitro has remained impossible with models either representing the outer membrane (OM) or the inner membrane (IM) solely. For the study of trans-envelope systems, however, no in vitro system is readily available. This therefore limits the study of transport processes between membranes, in particular the study of the biogenic transport processes responsible for OM formation. These processes utilise sophisticated and complex multi-protein machineries that have components in both membranes and in some cases span and tether the membranes together. One such system, the PqiABC complex, implicated in phospholipid transport, spans the periplasm with components in both the inner and outer membranes, tethering the two membranes together. Here we request rapid access to complete our studies utilising this trans-envelope complex as a scaffold to create a planar double membrane architecture of the gram-negative membrane on top of a solid support, providing a unique gram-negative envelope mimetic to probe envelope biology, significantly increasing the sophistication and accuracy of envelope based in vitro studies.