19F-NMR Reveals the Role of Mobile Loops in Product and Inhibitor Binding by the São Paulo Metallo-β-Lactamase

Martine I. Abboud, Philip Hinchliffe, Jgrgen Brem, Robert Macsics, Inga Pfeffer, Anne Makena, Klaus-Daniel Umland, Anna M Rydzik, Guo-Bo Li, James Spencer, Timothy D W Claridge, Christopher J Schofield

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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.
Original languageEnglish
Pages (from-to)3862–3866
Number of pages5
JournalAngewandte Chemie - International Edition
Volume56
Issue number14
Early online date2 Mar 2017
DOIs
Publication statusPublished - 22 Mar 2017

Keywords

  • antibiotic resistance
  • β-lactamases
  • NMR spectroscopy
  • protein structures
  • São Paulo metallo-β-lactamase

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    Abboud, M. I., Hinchliffe, P., Brem, J., Macsics, R., Pfeffer, I., Makena, A., Umland, K-D., Rydzik, A. M., Li, G-B., Spencer, J., Claridge, T. D. W., & Schofield, C. J. (2017). 19F-NMR Reveals the Role of Mobile Loops in Product and Inhibitor Binding by the São Paulo Metallo-β-Lactamase. Angewandte Chemie - International Edition, 56(14), 3862–3866. https://doi.org/10.1002/anie.201612185