GOURLEY DG, ALI ST, BOTTOMLEY JR, GILL AE, GRANT RM, LESLIE BW, RIDDELL MP, TAYLOR K, TYLER PD; Interscience Conference on Antimicrobial Agents and Chemotherapy (43rd: 2003: Chicago, Ill.).
Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 2003 Sep 14-17; 43: abstract no. F-345.
PanTherix Ltd., Glasgow, United Kingdom.
BACKGROUND: The mevalonate pathway is required for isoprenoid biosynthesis in the gram-positive pathogens Staphylococcus aureus, Streptococcus pneumoniae and Enterococcus faecalis, and is essential for bacterial viability. The pathway is also present in eukaryotes and, in man, the key enzyme of this pathway, HMG-CoA reductase, is the target of the statin class of cholesterol-lowering drugs. Statins are, however, very poor inhibitors of the bacterial enzymes. Human and bacterial HMG-CoA reductases are sufficiently divergent to allow development of selective inhibitors of the bacterial enzymes. A selective inhibitor would target the major gram-positive pathogens, including multi-drug resistant MRSA and VRE. Our aim is to exploit structural information to design such a selective inhibitor for antibacterial use. METHODS: HMG-CoA reductase genes from S. aureus (mvaA) and S. pneumoniae (mvaS) were cloned, and the enzymes over-expressed and purified. The kinetics and mechanism of these enzymes were studied using a previously developed assay. The protein from S. pneumoniae was crystallised and its structure determined by X-ray crystallography. The resulting high-resolution structure was then used for virtual screening in order to identify selective inhibitors. RESULTS: HMG-CoA reductases from S. aureus and S. pneumoniae were purified to homogeneity and characterized. Protein crystals of the S. pneumoniae enzyme were grown and the X-ray structure solved to high resolution. CONCLUSIONS: We have generated the first crystal structure of HMG-CoA reductase from a pathogenic bacterium. This structure, together with the structure of the human enzyme, affords an excellent opportunity to develop selective inhibitors of the bacterial enzyme.
Publication Types:
Keywords:
- Anti-Infective Agents
- Crystallography, X-Ray
- Enterococcus faecalis
- Humans
- Hydroxymethylglutaryl CoA Reductases
- Hydroxymethylglutaryl-CoA Reductase Inhibitors
- Hydroxymethylglutaryl-CoA Synthase
- Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent
- Kinetics
- Male
- Staphylococcal Infections
- Staphylococcus aureus
- Streptococcus pneumoniae
- drug therapy
- enzymology
Other ID:
UI: 102266353
From Meeting Abstracts