title: Active-site mutants of class B beta-lactamases: substrate binding and mechanistic study.
abstract: Increased resistance to beta-lactam antibiotics is mainly due to beta-lactamases. X-ray structures of zinc beta-lactamases unraveled the coordination of the metal ions, but their mode of action remains unclear. Recently, enzymes in which one of the zinc ligands was mutated have been characterized and their catalytic activity against several beta-lactam antibiotics measured. A molecular modeling study of these enzymes was performed here to explain the catalytic activity of the mutants. Coordination around the zinc ions influences the way the tetrahedral intermediate is bound; any modification influences the first recognition of the substrate by the enzyme. For all the studied mutants, at least one of the interactions fails, inducing a loss of catalytic efficiency compared to the wild type. The present studies show that the enzyme cavity is a structure of high plasticity both structurally and mechanistically and that local modifications may propagate its effects far from the mutated amino acid.
authors.affiliation: Service de Physique Expérimentale, Institut de Physique (B5), University of Liège, Sart-Tilman, 4000 Liège, Belgium. christelle.prosperi@ulg.ac.be
authors.forename: C
authors.initials: C
authors.lastname: Prosperi-Meys
authors.forename: D
authors.initials: D
authors.lastname: de Seny
authors.forename: G
authors.initials: G
authors.lastname: Llabres
authors.forename: M
authors.initials: M
authors.lastname: Galleni
authors.forename: J
authors.initials: J
authors.lastname: Lamotte-Brasseur
mesh_terms: D001409:Bacillus cereus / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D006639:Histidine / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009154:Mutation; D010400:Penicillin G / Q000378:metabolism; D010406:Penicillins / Q000378:metabolism; D011485:Protein Binding; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D010406:Penicillins; D006639:Histidine; D001618:beta-Lactamases; D003545:Cysteine; D010400:Penicillin G