title: Studying the active-site loop movement of the São Paolo metallo-β-lactamase-1†Electronic supplementary information (ESI) available: Procedures for protein expression and purification,19F-labelling, crystallisation, data collection, and structure determination, table of crystallographic data, table of crystallographic parameters and refinement statistics, figures showing binding mode and distances, procedures for mass spectrometry measurements, differential scanning fluorimetry measurements, stopped-flow measurements and other kinetics measurements. See DOI: 10.1039/c4sc01752hClick here for additional data file.
abstract: Metallo-β-lactamases (MBLs) catalyse the hydrolysis of almost all β-lactam antibiotics. We report biophysical and kinetic studies on the São Paulo MBL (SPM-1), which reveal its Zn(ii) ion usage and mechanism as characteristic of the clinically important di-Zn(ii) dependent B1 MBL subfamily. Biophysical analyses employing crystallography, dynamic19F NMR and ion mobility mass spectrometry, however, reveal that SPM-1 possesses loop and mobile element regions characteristic of the B2 MBLs. These include a mobile α3 region which is important in catalysis and determining inhibitor selectivity. SPM-1 thus appears to be a hybrid B1/B2 MBL. The results have implications for MBL evolution and inhibitor design.
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Brem
authors.affiliation: Department of Chemistry , Physical and Theoretical Chemistry Laboratory , University of Oxford , South Parks Road , Oxford , OX1 3QZ , UK . Email: justin.benesch@chem.ox.ac.uk.
authors.forename: Weston B
authors.initials: WB
authors.lastname: Struwe
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Rydzik
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Hanna
authors.initials: H
authors.lastname: Tarhonskaya
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Inga
authors.initials: I
authors.lastname: Pfeffer
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Emily
authors.initials: E
authors.lastname: Flashman
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Sander S
authors.initials: SS
authors.lastname: van Berkel
authors.affiliation: School of Cellular and Molecular Medicine , University of Bristol , Medical Sciences Building , Bristol , BS8 1TD , UK.
authors.forename: James
authors.initials: J
authors.lastname: Spencer
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Timothy D W
authors.initials: TD
authors.lastname: Claridge
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Michael A
authors.initials: MA
authors.lastname: McDonough
authors.affiliation: Department of Chemistry , Physical and Theoretical Chemistry Laboratory , University of Oxford , South Parks Road , Oxford , OX1 3QZ , UK . Email: justin.benesch@chem.ox.ac.uk.
authors.forename: Justin L P
authors.initials: JL
authors.lastname: Benesch
authors.affiliation: Department of Chemistry , University of Oxford , 12 Mansfield Road , Oxford , OX1 3TA , UK . Email: christopher.schofield@chem.ox.ac.uk.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Schofield