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

Journal article

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.

Authors: Abboud, M; Hinchliffe, P; Brem, J; Macsics, R; Pfeffer, I

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Wiley
• Journal: Angewandte Chemie International Edition
• Volume: 56
• Issue: 14
• Pages: 3862–3866
• Publication date: 2017-03-02
• Acceptance date: 2017-01-31
• DOI: 10.1002/anie.201612185
• EISSN: 1521-3757
• ISSN: 1433-7851
• Language: English
• Keywords: protein-observe 19F-NMR; São Paolo metallo-β-lactamase SPM-1 inhibition; Pseudomonas aeruginosa; metallo-β-lactamases MBL; antibacterial/antibiotic/β-lactam/carbapenem resistance