A study of outer membrane biogenesis in E. coli

The outer membrane (OM) of Escherichia coli is an essential organelle. The OM allows E. coli to interact with its environment and has a critical function as a barrier to prevent the entry of toxic molecules into the cell. The OM is composed of phospholipids, lipoproteins, outer membrane \\(\\beta\\)-barrel proteins (OMPs) and lipopolysaccharide (LPS). The correct ratio of these components is needed to ensure proper OM barrier function is maintained. Assembly of OMPs is performed by the Bam (\\(\\beta\\)-barrel assembly machinery) complex, lipoproteins by the Lol (Localisation of lipoproteins) pathway and LPS by the Lpt (LPS transport) pathway. The factors responsible for the assembly of phospholipids at the OM are unknown. \n \nThis study presents two key areas in understanding OM biogenesis. Firstly, a comprehensive mutagenesis screen was performed on the Bam complex member BamE. This analysis along with the structure of BamE has indicated crucial regions for BamE function. Secondly, we have performed a structure and function analysis on the previously uncharacterised protein, YraP. The structure of YraP has been solved and represents a novel fold. Additionally, we have obtained some functional evidence that suggest that YraP is involved in phospholipid biogenesis in the OM.