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Title: Application of Structural Genomics for Developing Novel Antibacterial Agents
Author: Das, Sanjan Kumar
ISNI:       0000 0001 3405 2077
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2007
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The structure-based design of novel antibiotics has great potentiaL To achieve this goal the first step is to gather structural information on relevant proteins and utilise this to find inhibitors through a process of screening and subsequent optimising using structural data. In this· pilot study a number of essential gene products from Bacillus subtilis were targeted for structural studies where homologues were present in pathogenic microorganisms. In the initial analysis 41 essential genes of unknown structure or function were selected for structure determination. From this list 9 genes were rejected as their protein products were assigned as possible membrane proteins by hydropathy analysis. From the remaining 32 genes on the list 12 (ysxC, thiD, ywaF, ydiC, ydiE, yrrA, yabH, ylxR, yqiB, yacN, ymjL and luxS) were selected for study as part of this thesis. Recombinant DNA technology was used to produce pure protein for crystallisation trials and subsequent structure determination by X-ray crystallography. Successful overexpression was achieved for 8 of the protein targets ofwhich 7 were found to be soluble. 6 ofthese were taken forward for crystallisation as part of this work with the 7th being·pursued by another member of the laboratory due to project overlap. Of these 6 proteins 5 were successfully purified and crystallised (LuxS, YsxC, YacN, YabH, and ThiD) and structures were determined for all but ThiD. The successful crystallisation of all of the 5 soluble proteins was unexpected and maybe related to the characteristics of the structure determination pipeline which requires the expression of the protein product in a soluble form and at high leveL This may result in the selection of proteins with a higher tendency to crystallise. The gene to structure conversion rate'obtained here (33%) is equivalent to that obtained for the overall programme on all 32 genes (34%) suggesting that 4 bacterial proteins 1/3rd of the proteome can be analysed as part of a structural genomics programme.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available