Exploring the multiple applications of the membrane-disrupting Styrene Maleic Acid

Harris, Craig (2017). Exploring the multiple applications of the membrane-disrupting Styrene Maleic Acid. University of Birmingham. M.Phil.

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Abstract

Poly(styrene-co-maleic) acid (SMA) has demonstrated great potential as an interrogation tool for cell membranes. SMA can insert into the cell membrane excising discs of lipid and any associated membrane proteins. The potential applications of SMA in membrane protein study and the specific release of the periplasmic fraction of E. coli were investigated in this project. There is a well-documented disparity in the number of structurally characterised soluble proteins versus membrane proteins due to a number of factors. Whilst solubilisation and subsequent characterisation of membrane proteins has been achieved, lack of a universal membrane solubilisation protocol is restricting the growth of membrane protein study. SMA was applied to three membrane proteins (FtsA, MurJ-mfGFP and FtsW-mfGFP) leading to their solubilisation and purification with conventional methods. FtsA and MurJ-mfGFP were also characterised with CD and svAUC.
Periplasmic targeting of recombinant proteins in E. coli offers many advantages over cytoplasmic production. Existing methods for selective periplasmic extraction are expensive, time consuming and inefficient In this report we investigated whether SMA can disrupt the E. coli outer membrane to selectively release the periplasmic contents. We discovered that SMA performed favorably compared to an existing method at selectively releasing two periplasm-targeted proteins.

Type of Work:

Thesis (Masters by Research > M.Phil.)

Award Type:

Masters by Research > M.Phil.

Supervisor(s):