Oxidative protein folding is an important process for the function of many proteins, which includes both folding and formation of native disulphide bonds. Oxidative protein folding of disulphide bond containing proteins is challenging and often a key problem of recombinant protein production in vitro, as disulphide bond formation is catalysed by various oxidoreductase systems in cells. The Mia40-Erv1 system is a sulfhydryl oxidoreductase system that catalyses oxidative folding of the newly imported proteins in the mitochondrial intermembrane space. We have carried out many studies in understanding the structure and function of Mia40 and Erv1, and established good systems for the purification and characterisation of these proteins. In this study, the effects of a recombinant Mia40-Erv1 system on oxidative folding of industrial and therapeutically important high-value production proteins (e.g. TGF-b1, TGF-b3) will be investigated, and the Mia40-Erv1 system will be optimised to enhance the oxidative folding of the target proteins. The key techniques required for this project include protein purification from E. coli., protein folding and functional characterisation using electrophoresis, chromatography, and various spectroscopic methods. Therefore, this project will provide an excellent opportunity for a self-motivated student to learn and develop many essential skills in protein studies. The project is suitable for both MPhil and PhD studies.
Entry Requirements:
Candidates are expected to hold (or about to obtain) a minimum second class (for MPhil study) and upper second class (for PhD study) honours degree (or equivalent) in biochemistry, cell biology, chemistry or a related subject. Candidates with experience in protein folding study or with an interest in protein folding and protein biotechnology are encouraged to apply.
For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (
https://www.bmh.manchester.ac.uk/study/research/apply/). Informal enquiries may be made directly to the primary supervisor.
For international students we also offer a unique 4 year PhD programme that gives you the opportunity to undertake an accredited Teaching Certificate whilst carrying out an independent research project across a range of biological, medical and health sciences. For more information please visit
http://www.internationalphd.manchester.ac.uk
Funding Notes
This project is offered as a one year MPhil or a three year PhD. Applications are invited from self-funded students. This project has a Band 2 fee. Details of our different fee bands can be found on our website (View Website).
As an equal opportunities institution we welcome applicants from all sections of the community regardless of gender, ethnicity, disability, sexual orientation and transgender status. All appointments are made on merit.
References
Ang SK, & Lu H. (2009) Deciphering structural and functional roles of individual disulfide bonds of the mitochondrial sulfhydryl oxidase Erv1p, J Biol Chem. 284, 28754-61.
Gisby MF, Mellors P, et al, & Day A (2011) A synthetic gene increases TGFβ3 accumulation by 75-fold in tobacco chloroplasts enabling rapid purification and folding into a biologically active molecule. Plant Biotechol J 9, 618-628.
Hell K (2008) The Erv1-Mia40 disulfide relay system in the intermembrane space of mitochondria, Biochim Biophys Acta. 1783, 601-9. Review
Riemer J, Bulleid N & Herrmann JM (2009) Disulfide formation in the ER and mitochondria: two solutions to a common process, Science. 324,1284-7. Review
Spiller MP, Ang SK et al & Lu H (2013) Identification and characterization of mitochondrial Mia40 as an iron-sulfur protein. Biochem J. 455, 27-35.
