Understanding bacterial cell division proteins using novel nanoencapsulation methods

Stroud, Zoe (2020). Understanding bacterial cell division proteins using novel nanoencapsulation methods. University of Birmingham. Ph.D.

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Abstract

The proteins of the Escherichia coli divisome are essential for bacterial viability, and have attracted increasing interest as potential targets for new antibiotics. Despite this, very little is known about the structure and function of the FtsBLQ divisomal complex, due to the complexities of studying integral membrane proteins. The development of the use of styrene-maleic acid (SMA) as a solubilisation technique over the past decade offers an alternative strategy for the solubilisation of the complex. SMA solubilises proteins with intact transmembrane regions and the surrounding native lipid environment.

This thesis aimed to establish whether SMA is an effective method for the solubilisation of the FtsBLQ complex. Using SMA, the individual proteins of the FtsBLQ complex were solubilised successfully and characterised by a range of methods. Additionally, the FtsB/FtsL subcomplex was co-expressed and purified, and a model of the association of these proteins was supported with biophysical studies. These results demonstrate that SMA supports the solubilisation of divisomal proteins in the native lipid environment and maintains protein-protein interactions. The application of analytical ultracentrifugation to SMA-solubilised proteins was also investigated, demonstrating the applicability of SMA to a wide range of downstream techniques.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):