Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.661088
Title: Structural and functional studies of Drosophila alcohol dehydrogenase
Author: Ribas de Pouplana, L.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1993
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Abstract:
The enzyme alcohol dehydrogenase of the fruit-fly Drosophila (DADH) catalyzes the same reaction as mammalian alcohol dehydrogenases, transforming alcohols into aldehydes through the reduction of nicotinamide-adenine-dinucleotide. Despite this apparently identical catalytic behaviour the enzyme has a strikingly different three-dimensional architecture to the mammalian alcohol dehydrogenases. The study of the structure-function relationships of DADH is of interest because large amounts of biochemical, evolutionary and genetical data have accumulated which require structural information on the enzyme for its proper interpretation. The aim of this project was two-fold: to set up a suitable system for protein engineering studies on DADH and to start such studies by producing and analyzing a first set of site-directed mutants. The first part of the project involved: a) creating suitable genetic vectors for the introduction of mutations, their sequencing and the expression of the mutated enzymes in yeast; b) the development of a purification method to obtain pure enzyme solutions from the expressing yeast culture; c) the development of biochemical and biophysical assays for the evaluation of the effects of the mutations and, d) the construction of a three-dimensional model for the enzyme that could offer structural explanations to such effects, given that no three-dimensional structure is as yet available. The second part consisted of the actual introduction of five different point mutations, designed to test the structure of the putative cofactor binding site, and their further evaluation using the system set up in the first place.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.661088  DOI: Not available
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