Caractérisation structurale et étude de l'inhibityion de la 1-déoxy-D-xylulose-5-phosphate réductoisomérase (DXR) de E. coli et de la DXR-like (DRL) de B. abortus, par des analogues de la fosmidomycine

Résumé

The 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has come under increased scrutiny as a target for novel antimalarial, antibacterial and antituberculosis agents. 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) is a key enzyme of the pathway that catalyzes the rearrangement and nicotinamide adenine dinucleotide phosphate (NADPH)-dependent reduction of 1-deoxy-D-xylulose 5-phosphate (DXP) to MEP. The unique properties of DXR make it a remarkable and rational target for drug design. First, it is a vital enzyme for synthesis of isoprenoids in algae, plants, several eubacteria including the pathogenic bacteria like Bacillus anthracis, Helicobacter pylori, Yersinia pestis, Mycobacterium tuberculosis and the malarial parasite, Plasmodium falciparum. Second, there are no functional equivalents to DXR in humans, making it an attractive target for therapeutic intervention. Third, DXR appears to be a valid target and the results from fosmidomycin, the only available DXR inhibitor under clinical trials, suggests synergistic effects with the lincosamide antibiotics, lincomycin and clindamycin. Despite drug design efforts in this area, no successful drug specifically designed to inhibit DXR has emerged yet. Recently, it has been shown that Fosmidomycin inhibited growth of B. abortus cells expressing the Escherichia coli GlpT transporter (required for fosmidomycin uptake), confirming that a DXR-like (DRL) activity exists in these bacteria. The B. abortus DRL protein was found to belong to a family of uncharacterized proteins similar to homoserine dehydrogenase. Our work was focus on the production, purification and characterization of the E. coli DXR enzyme and the B. abortus DRL enzyme. This allowed us to study three series of analogs of fosmidomycin, a specific competitive inhibitor of DXR, available through collaboration with the University of Gent (Prof S.VanCalenberg). Two approaches were used to characterize interaction between inhibitor and enzyme, a theoretical approach by docking and an experimental approach by crystallography.

la date de réponse	3 oct. 2014
langue originale	Français
L'institution diplômante	Universite de Namur
Superviseur	Johan Wouters (Promoteur), Carmela Aprile (Président), Xavier De Bolle (Jury), Steve Lanners (Jury), Laurence Leherte (Jury) & Paulette Charlier (Jury)