Objective To characterise the enzymatic activities of the catalase-peroxidase from Mycobacterium tuberculosis and related variants . To characterise the molecular mechanism and putative intermediates associated with isoniazid activation . To determine the structure of the active site of catalase-peroxidase using NMR and/or x-ray crystallographic methods . To prepare synthetic lead compounds targeted for drug-resistant strainsTo develop a new line of therapeutics aimed at providing an alternative treatment regime for multi-drug resistant strains of Mycobacterium tuberculosis, we take advantage of the participants' expertise in biochemistry, organic synthesis, NMR and crystallography. A functional description of the active site or sites of the recombinant katG gene product, catalase-peroxidase, will be established using a variety of techniques including visible spectrophotometry, electron paramagnetic resonance and nuclear magnetic resonance spectroscopy. Hypothetical mechanisms for the activation of the anti-tuberculosis drug by this enzyme will be tested using synthesised "intermediates," in both biological and model chemical (enzyme-free) systems. Classical analytical chemistry methods including NMR, HPLC, mass spectrometry and stopped-flow kinetics will be integrated with genetic engineering and other biologically based-studies to observe and characterise critical intermediates in the reaction pathway. Parallel efforts to obtain additional information about active site structure and ligand interactions will be carried out using paramagnetic NMR and x-ray crystallographic methods. Comparison of the functional and structural data will contribute insight into the metabolic activation mechanism of isoniazid, suggesting new lead compounds for alternative treatment regimes. Fields of science natural sciencesearth and related environmental sciencesgeologymineralogycrystallographymedical and health sciencesclinical medicinepneumologytuberculosismedical and health sciencesbasic medicinepharmacology and pharmacydrug resistancemultidrug resistancenatural scienceschemical sciencesanalytical chemistrymass spectrometrynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes Programme(s) FP4-BIOMED 2 - Specific research, technological development and demonstration programme in the field of biomedicine and health, 1994-1998 Topic(s) 1.1 - Pharmacotoxicology Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator Imperial College of Science Technology and Medicine EU contribution No data Address Exhibition Road SW7 2AY LONDON United Kingdom See on map Total cost No data Participants (2) Sort alphabetically Sort by EU Contribution Expand all Collapse all Università degli Studi di Firenze Italy EU contribution No data Address Via Gino Capponi 9 50121 Firenze See on map Total cost No data Université Henri Poincaré Nancy 1 France EU contribution No data Address BLD. DES AIGUILLETTES 54506 VANDOEUVRE LES NANCY See on map Total cost No data
