Supported lipid bilayers (SLBs) for E. coli proteins study

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Daniela Garcia, Ariadna Martos, Petra Schwille


The study of in vivo cellular events is not always feasible due to the high complexity of living organisms. Even though a full understanding of a system is the final goal, sometimes one must focus in the minimal set of components that carry on a specific cellular event in vitro. One particular process that has been studied by many years is cell division. In E. coli, the set of proteins involved in the process have been identified, but the detailed performance of each component in the whole event is not yet understood. Most of the proteins somehow interact with the cell membrane, thus, when reconstructing the minimal set of players for cell division, model cell membranes are an important element to take into account. In recent years, different model lipid bilayers have been described with particular properties, complexities and methods of preparation. Supported lipid bilayers (SBLs) are lipid bilayer structures on a solid support; they are stable and durable models suitable for high resolution imaging characterization. SBLs flexibility allows the use of different lipids composition to emulate the native components present in a specific cell membrane. In our case of interest, E.coli membrane is primarily composed by phosphatidylglycerol, phosphatidylethanolamine, cardiolipin, however, it also presents a high percent of membrane proteins. The main goal of this work is to develop different SLBs for the study of the bacterial cell division machinery. In the first place SLBs with different lipid composition were tested, trying to obtain minimal sets of mixtures that are able to support the performance of the whole cell-division machinery. In second place, SBLs that resembles the high protein content of E. coli membrane are sought, either by using E. coli lipid extracts doped either with ZipA proteoliposomes or, inner membrane vesicles (IMVs).