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

Harris, Craig (2017)
M.Phil. thesis, University of Birmingham.

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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:M.Phil. thesis.
Supervisor(s):Dafforn, Timothy
School/Faculty:Colleges (2008 onwards) > College of Life & Environmental Sciences
Department:School of Biosciences
Subjects:QH301 Biology
QR Microbiology
Institution:University of Birmingham
ID Code:7276
This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder.
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