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19F-NMR reveals the role of mobile loops in product and inhibitor binding by the São Paulo metallo-β-lactamase

DOI: 10.1002/anie.201612185 DOI Help

Authors: Martine I. Abboud (University of Oxford) , Philip Hinchliffe (University of Bristol) , Jurgen Brem (University of Oxford) , Robert Macsics (University of Oxford) , Inga Pfeffer (University of Oxford) , Anne Makena (University of Oxford) , Klaus-Daniel Umland (University of Oxford) , Anna M. Rydzik (University of Oxford) , Guo-Bo Li (University of Oxford) , James Spencer (University of Bristol) , Timothy D. W. Claridge (University of Oxford) , Christopher J. Schofield (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Angewandte Chemie International Edition , VOL 5

State: Published (Approved)
Published: March 2017
Diamond Proposal Number(s): 8922

Open Access Open Access

Abstract: Resistance to β-lactam antibiotics mediated by metallo-β-lactamases (MBLs) is a growing problem. We describe the use of protein-observe 19F-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-ZnII active site, were selectively 19F-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.

Journal Keywords: antibiotic resistance; β-lactamases; NMR spectroscopy; protein structures; São Paulo metallo-β-lactamase

Diamond Keywords: Bacteria

Subject Areas: Chemistry, Biology and Bio-materials, Medicine

Instruments: I24-Microfocus Macromolecular Crystallography

Added On: 13/03/2017 11:59


Discipline Tags:

Pathogens Antibiotic Resistance Infectious Diseases Health & Wellbeing Biochemistry Chemistry Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)