title: 19F-NMR Reveals the Role of Mobile Loops in Product and Inhibitor Binding by the São Paulo Metallo-β-Lactamase.
abstract: Resistance to β-lactam antibiotics mediated by metallo-β-lactamases (MBLs) is a growing problem. We describe the use of protein-observe19F-NMR (PrOF NMR) to study the dynamics of the São Paulo MBL (SPM-1) from β-lactam-resistant Pseudomonas aeruginosa. Cysteinyl variants on the α3 and L3 regions, which flank the di-ZnIIactive site, were selectively19F-labeled using 3-bromo-1,1,1-trifluoroacetone. The PrOF NMR results reveal roles for the mobile α3 and L3 regions in the binding of both inhibitors and hydrolyzed β-lactam products to SPM-1. These results have implications for the mechanisms and inhibition of MBLs by β-lactams and non-β-lactams and illustrate the utility of PrOF NMR for efficiently analyzing metal chelation, identifying new binding modes, and studying protein binding from a mixture of equilibrating isomers.
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Martine I
authors.initials: MI
authors.lastname: Abboud
authors.affiliation: School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK.
authors.forename: Philip
authors.initials: P
authors.lastname: Hinchliffe
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Brem
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Robert
authors.initials: R
authors.lastname: Macsics
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Inga
authors.initials: I
authors.lastname: Pfeffer
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Makena
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Klaus-Daniel
authors.initials: KD
authors.lastname: Umland
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Rydzik
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Guo-Bo
authors.initials: GB
authors.lastname: Li
authors.affiliation: School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK.
authors.forename: James
authors.initials: J
authors.lastname: Spencer
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Timothy D W
authors.initials: TD
authors.lastname: Claridge
authors.affiliation: Department of Chemistry, University of Oxford, 12 Mansfield Road, OX1 3TA, Oxford, UK.
authors.forename: Christopher J
authors.identifier: 0000-0002-0290-6565
authors.initials: CJ
authors.lastname: Schofield
mesh_terms: D001665:Binding Sites / Q000187:drug effects; D066235:Fluorine-19 Magnetic Resonance Imaging; D008958:Models, Molecular; D008968:Molecular Conformation; D065093:beta-Lactamase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001618:beta-Lactamases / Q000378:metabolism*
chemical_list: D065093:beta-Lactamase Inhibitors; D001618:beta-Lactamases
keywords: NMR spectroscopy; São Paulo metallo-β-lactamase; antibiotic resistance; protein structures; β-lactamases