Understanding membrane protein biogenesis is of central importance for modern society as membrane proteins are not only crucial for transport or acting as receptors, they also comprise a major drug target for phamaceutical industry. In E. coli membrane protein biogenesis requires components of the well- described SRP and Sec pathways for targeting to and insertion into the membrane. However, a clear picture of the integration of membrane proteins into the lipid bilayer has not yet emerged. Recently, a membrane protein (YidC) essential for viability of E. coli has been identified. Members of the YidC family exist in all three domains of life where they have multiple functions in the integration and assembly of a large variety of membrane protein complexes.
The host institution has established an efficient expression and purification protocol for YidC which allows now for the detailed structural and biochemical characterization of YidC alone as well as in complex with functionally relevant partners. Several detergents will be systematically screened for their influence on the oligomeric state of YidC as well as on its function. The interactions of YidC with the SRP system will be analysed and potential binding partners e.g. the SRP receptor, will be included in t he biochemical and structural studies. Besides E. coli, thermophilic bacteria will be used as source for YidC and also chimeras will be tested for structure analysis.
The results obtained will not only contribute to the molecular details of membrane protein biogenesis, but will also add to a rational optimisation of membrane protein production by engineering the membrane insertion pathway. Due to the combination of biochemical and biophysical methods this project represents an excellent opportunity to strengthen my background in membrane protein crystallography and to complement my previous expertise with methods from molecular biology, biochemistry of protein complexes and electron microscopy.
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