An Asymmetric Protein-Containing Gram-Negative bacterial Outer membrane Model

We have been developing advanced planar models of the Gram negative bacterial outer membrane as precise and accurate structural assay systems for antibiotic testing by neutron reflectometry. Here we wish to advance the accuracy and utility of these systems by incorporating integral membrane proteins into the models in the form of porins and depositing the asymmetric membranes through a two stage directed in-situ self-assembly methodology, removing the need for time consuming layer-by-layer deposition. Porin : phospholipid vesicles will be ruptured onto silicon surfaces to produce protein-phospholipid bilayers. Lipopolysaccharides will then be introduced into the bulk media which will exchange the outer leaflet lipids forming a porin:phospholipid:lipopolysaccharide membrane. We intend to control the asymmetry of this construct through bridging the inner leaflet phospholipids to the SiO2 layer based on previous observations by us. Finally we will test the stability of the sample under serum solution and temperature conditions. If successful this study will pave the way for a new level of accuracy in interfacial biomimetics using a much simplified fabrication route. This is a resubmission taking into account the panels previous comments by providing preliminary data in the proposal.