Figure 1. Colicin translocation across the outer and inner membrane and killing mechanism. Colicins recognize outer membrane proteins on the bacterial surface, which they use to cross the outer membrane. From there, they can directly exert their killing mechanism on the inner membrane by forming pores, or transport themselves across the inner membrane and act as nucleases within the cytoplasm
A second highly relevant part of this project will be to measure the conductance of these mutants, as well as other OmpX hybrid mutants, at the single-molecule level, to better understand how these mutants affect the transport properties of OmpX embedded in a lipid membrane. This can help us to understand potential structural differences between the various OmpX mutants, as well as give insight into the evolution of OMP β-barrels and their transport properties. These measurements will be carried out in close-collaboration with U.S. partners, who will use the experimental information from the conductance measurements to carry out in silico modeling of how ion transport through OmpX occurs.
This is a highly interdisciplinary and collaborative project. The student will be closely supervised by PhDs and Postdocs currently in the group, and will be trained in all methods necessary to complete the work. Methods to be learned will include:
- Molecular biology and cloning
- Large scale membrane protein expression and purification
- In vivo and in vitro protein folding and functional assay design
- Single-molecule conductance measurements
MSc candidates for this project are expected to bring good theoretical knowledge in biochemistry and molecular biology