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Pellecchia, Maurizio; Meininger, David; Dong, Qing; Chang, Edcon; Jack, R M; Sem, Daniel S.

doi:10.1023/a:1014256707875

Cited by 100 publications

(87 citation statements)

References 15 publications

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“…Although the fluorescence-based assay is a robust technique to search for very potent inhibitors, it becomes more ambiguous in detecting weaker ligands (Ͼ100 M), possibly due to interference introduced by test compounds (normally used at high concentration) in the spectrophotometric assay. For this reason, we relied on a NMR-based enzymatic assay, which is unlikely to lead to false positives (16)(17)(18)(19)(20)(21)(22)(23). Recently, the use of 19 F-1D NMR to detect enzyme activity and inhibition both in proteases and kinases has been reported (22).…”

Section: Resultsmentioning

confidence: 99%

Efficient synthetic inhibitors of anthrax lethal factor

Forino

Johnson

Wong

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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125

144

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Inhalation anthrax is a deadly disease for which there is currently no effective treatment. Bacillus anthracis lethal factor (LF) metalloproteinase is an integral component of the tripartite anthrax lethal toxin that is essential for the onset and progression of anthrax. We report here on a fragment-based approach that allowed us to develop inhibitors of LF. The small-molecule inhibitors we have designed, synthesized, and tested are highly potent and selective against LF in both in vitro tests and cell-based assays. These inhibitors do not affect the prototype human metalloproteinases that are structurally similar to LF. Initial in vivo evaluation of postexposure efficacy of our inhibitors combined with antibiotic ciprofloxican against B. anthracis resulted in significant protection. Our data strongly indicate that the scaffold of inhibitors we have identified is the foundation for the development of novel, safe, and effective emergency therapy of postexposure inhalation anthrax.

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Section: Resultsmentioning

confidence: 99%

Efficient synthetic inhibitors of anthrax lethal factor

Forino

Johnson

Wong

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

125

144

View full text Add to dashboard Cite

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“…We therefore conclude that the catalytic loop and the nicotinamide moiety of NADPH are simultaneously required for their correct positioning at the active site. A recent NMR study demonstrated the proximity of a methionine and an isoleucine residue to the nicotinamide ring portion of NADPH (41). Potential candidates at the active site are Met-214 or Met-276 and Ile-13 or Ile-218, respectively, which all represent almost strictly conserved residues.…”

Section: Binding Of Manganese and Fosmidomycin And Implicationsmentioning

confidence: 99%

Structural Basis of Fosmidomycin Action Revealed by the Complex with 2-C-Methyl-d-erythritol 4-phosphate Synthase (IspC)

Steinbacher

Kaiser

Eisenreich

et al. 2003

Journal of Biological Chemistry

171

184

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2-C-Methyl-D-erythritol 4-phosphate synthase (IspC) is the first enzyme committed to isoprenoid biosynthesis in the methylerythritol phosphate pathway, which represents an alternative route to the classical mevalonate pathway. As it is present in many pathogens and plants, but not in man, this pathway has attracted considerable interest as a target for novel antibiotics and herbicides. Fosmidomycin represents a specific high-affinity inhibitor of IspC. Very recently, its anti-malaria activity in man has been demonstrated in clinical trials. Here, we present the crystal structure of Escherichia coli IspC in complex with manganese and fosmidomycin at 2.5 Å resolution. The (N-formyl-N-hydroxy)amino group provides two oxygen ligands to manganese that is present in a distorted octahedral coordination, whereas the phosphonate group is anchored in a specific pocket by numerous hydrogen bonds. Both sites are connected by a spacer of three methylene groups. The substrate molecule, 1-D-deoxyxylulose 5-phosphate, can be superimposed onto fosmidomycin, explaining the stereochemical course of the reaction.

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“…Selective-labeling approaches have been used to measure intermolecular NOEs without the need for explicit side-chain assignment. [6] Ligandbased techniques, such as TrNOE [7] or TrCCR, [8] define the structure of a ligand in its protein-bound state but do not give information about its binding mode. Sometimes they even require isotope labeling of the ligand.…”

Section: Introductionmentioning

confidence: 99%

How Much NMR Data Is Required To Determine a Protein–Ligand Complex Structure?

et al. 2005

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Here we present an NMR-based approach to solving protein-ligand structures. The procedure is guided by biophysical, biochemical, or knowledge-based data. The structures are mainly derived from ligand-induced chemical-shift perturbations (CSP) induced in the resonances of the protein and ligand-detected saturated transfer difference signals between ligands and selectively labeled proteins (SOS-NMR). Accuracy, as judged by comparison with X-ray results, depends on the nature and completeness of the experimental data. An experimental protocol is proposed that starts with calculations that make use of readily available chemical-shift perturbations as experimental constraints. If necessary, more sophisticated experimental results have to be added to improve the accuracy of the protein-ligand complex structure. The criteria for evaluation and selection of meaningful complex structures are discussed. These are exemplified for three complexes, and we show that the approach bridges the gap between theoretical docking approaches and complex NMR schemes for determining protein-ligand complexes; especially for relatively weak binders that do not lead to intermolecular NOEs.

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Untitled

Cited by 100 publications

References 15 publications

Efficient synthetic inhibitors of anthrax lethal factor

Efficient synthetic inhibitors of anthrax lethal factor

Structural Basis of Fosmidomycin Action Revealed by the Complex with 2-C-Methyl-d-erythritol 4-phosphate Synthase (IspC)

How Much NMR Data Is Required To Determine a Protein–Ligand Complex Structure?

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