Objetivo
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 strains
To 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.
Ámbito científico
- natural sciencesearth and related environmental sciencesgeologymineralogycrystallography
- medical and health sciencesclinical medicinepneumologytuberculosis
- medical and health sciencesbasic medicinepharmacology and pharmacydrug resistancemultidrug resistance
- natural scienceschemical sciencesanalytical chemistrymass spectrometry
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes
Tema(s)
Convocatoria de propuestas
Data not availableRégimen de financiación
CSC - Cost-sharing contractsCoordinador
SW7 2AY LONDON
Reino Unido