Crystal Structure of the IMP-1 Metallo β-Lactamase from Pseudomonas aeruginosa and Its Complex with a Mercaptocarboxylate Inhibitor: Binding Determinants of a Potent, Broad-Spectrum Inhibitor
| Autor: | Brian Peter SmithKline Beecham Pharm Clarke, Moreno Galleni, David J. Payne, Bateson John Hargreaves, Ceri Lewis, Christy A. Cheever, Sherin S. Abdel-Meguid, Pam Rowling, Nestor O. Concha, Stewart C. Pearson, Jean-Marie Frere, Cheryl A. Janson |
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| Rok vydání: | 2000 |
| Předmět: |
Models
Molecular Conformational change Protein Conformation Stereochemistry Molecular Sequence Data Static Electricity Glycine Tetrazoles Plasma protein binding Crystallography X-Ray Biochemistry beta-Lactamases Substrate Specificity Hydrolase Molecular replacement Enzyme Inhibitors Binding site Beta-Lactamase Inhibitors Conserved Sequence Binding Sites biology Lysine Water Active site Hydrogen Bonding Zinc Drug Design Pseudomonas aeruginosa biology.protein Oxyanion hole beta-Lactamase Inhibitors Protein Binding |
| Zdroj: | Biochemistry. 39:4288-4298 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi992569m |
| Popis: | Metallo beta-lactamase enzymes confer antibiotic resistance to bacteria by catalyzing the hydrolysis of beta-lactam antibiotics. This relatively new form of resistance is spreading unchallenged as there is a current lack of potent and selective inhibitors of metallo beta-lactamases. Reported here are the crystal structures of the native IMP-1 metallo beta-lactamase from Pseudomonas aeruginosa and its complex with a mercaptocarboxylate inhibitor, 2-[5-(1-tetrazolylmethyl)thien-3-yl]-N-[2-(mercaptomethyl)-4 -(phenylb utyrylglycine)]. The structures were determined by molecular replacement, and refined to 3.1 A (native) and 2.0 A (complex) resolution. Binding of the inhibitor in the active site induces a conformational change that results in closing of the flap and transforms the active site groove into a tunnel-shaped cavity enclosing 83% of the solvent accessible surface area of the inhibitor. The inhibitor binds in the active site through interactions with residues that are conserved among metallo beta-lactamases; the inhibitor's carboxylate group interacts with Lys161, and the main chain amide nitrogen of Asn167. In the "oxyanion hole", the amide carbonyl oxygen of the inhibitor interacts through a water molecule with the side chain of Asn167, the inhibitor's thiolate bridges the two Zn(II) ions in the active site displacing the bridging water, and the phenylbutyryl side chain binds in a hydrophobic pocket (S1) at the base of the flap. The flap is displaced 2.9 A compared to the unbound structure, allowing Trp28 to interact edge-to-face with the inhibitor's thiophene ring. The similarities between this inhibitor and the beta-lactam substrates suggest a mode of substrate binding and the role of the conserved residues in the active site. It appears that the metallo beta-lactamases bind their substrates by establishing a subset of binding interactions near the catalytic center with conserved characteristic chemical groups of the beta-lactam substrates. These interactions are complemented by additional nonspecific binding between the more variable groups in the substrates and the flexible flap. This unique mode of binding of the mercaptocarboxylate inhibitor in the enzyme active site provides a binding model for metallo beta-lactamase inhibition with utility for future drug design. |
| Databáze: | OpenAIRE |
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