ABCG2 is one of three human ATP binding cassette transporters that are functionally capable of exporting a diverse range of substrates from cells. The physiological consequence of ABCG2 multidrug transport activity in leukaemia, and some solid tumours is the acquisition of cancer multidrug resistance. The molecular basis for drug resistance is currently unknown, and we have scant information regarding regions on the protein responsible for drug and inhibitor interaction [1,2]. This project aims to address these questions.
In terms of drug binding it is known that 2 residues in the lower (cytoplasmic) region of TM helix 3 in ABCG2 are important in drug recognition. In the current proposal we would examine ABCG2 drug binding as follows:
i) Mutation of other residues in TM helix 3.
ii) Further characterization of residues of importance in TM helix 3
In part i) we would make mutations through the length of TM3 and then produce stable cell lines expressing these mutant forms. We would then confirm membrane localization of mutants, and then probe function using two routine functional assays (fluorescence based). This would enable us to identify residues that are possible part of a drug binding site on the protein, or residues which contribute to the known allostery between drug binding and ATP hydrolysis.
In part ii) we would take mutants of interest and examine them in an alternative experimental system that allows us to determine how ABCG2:drug interactions are communicated to the ATP hydrolysis of this pump. This would enable us to develop a molecular understanding of drug transport by this important multidrug pump.
Through the project the student would receive training in a number of different techniques including molecular biology, cell culture, fluorescence-based assays, confocal microscopy, membrane and protein biochemistry. The student would also receive training in data analysis, reading the scientific literature, and scientific writing and presentation. The student would contribute to a vibrant research group, and would make presentations to the group and the wider scientific community.
Home and EU applicants should contact the supervisor to determine the current funding status for this project. International applicants should visit our page for information regarding fees and funding at the University http://www.nottingham.ac.uk/studywithus/international-applicants/scholarships-fees-and-finance/scholarships/index.aspx
Wong, K., Ma, J., Rothnie, A., Biggin, P. C., and Kerr, I. D. (2014) Towards understanding promiscuity in multidrug efflux pumps, Trends in biochemical sciences 39, 8-16.
Kerr, I. D., Haider, A. J., and Gelissen, I. C. (2011) The ABCG family of membrane-associated transporters: you don't have to be big to be mighty, Br J Pharmacol 164, 1767-1779
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