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X-ray crystallographic studies of therapeutic enzymes: nitroreductase and AKR1C3

Lovering, Andrew Lee (2003)
Ph.D. thesis, University of Birmingham.

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The \(Escherichia\) \(coli\) enzyme nitroreductase has been proposed as a candidate for the Gene-Directed Enzyme Prodrug Therapy approach in treating cancer. Structural studies on the enzyme were instigated in a first step towards improving enzyme activity. The enzyme was crystallized with the substrate analogue, nicotinic acid, and the structures of three crystal forms obtained. The fold has a mixed a/P structure, with a molecule of nicotinic acid bound next to the FMN cofactor. Several active site residues were identified as candidates for mutation. This procedure produced many mutant enzymes with increased catalytic activity. One double and four single mutants were chosen from these and crystal structures determined. The resulting information from this, and the establishment of a proof of principle, provides the basis for iterative cycles of enzyme improvement.

The human hydroxysteroid dehydrogenase AKR1C3 has been proposed to play a role in prostaglandin metabolism. Its inhibition by non-steroidal anti-inflammatory drugs may be important in a tumour differentiation strategy. AKR1C3 was crystallized, and the structure solved with bound nucleotide cofactor and several inhibitors, including the drugs indomethacin and flufenamic acid. Having obtained information on drug binding to AKR1C3, selective inhibitors can be designed, avoiding inhibition of "housekeeping" enzymes such as cyclo-oxygenases.

Type of Work:Ph.D. thesis.
Supervisor(s):White, Scott and Hyde, Eva
School/Faculty:Schools (1998 to 2008) > School of Biosciences
Department:School of Biosciences
Subjects:QH301 Biology
RM Therapeutics. Pharmacology
Institution:University of Birmingham
Library Catalogue:Check for printed version of this thesis
ID Code:3588
This unpublished thesis/dissertation is copyright of the author and/or third parties.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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