19 F-NMR Reveals the Role of Mobile Loops in Product and Inhibitor Binding by the Sao Paulo Metallo-?-Lactamase.

ABSTRACT: Resistance to ?-lactam antibiotics mediated by metallo-?-lactamases (MBLs) is a growing problem. We describe the use of protein-observe 19 F-NMR (PrOF?NMR) to study the dynamics of the São Paulo MBL (SPM-1) from ?-lactam-resistant Pseudomonas aeruginosa. Cysteinyl variants on the ?3 and L3 regions, which flank the di-ZnII active site, were selectively 19 F-labeled using 3-bromo-1,1,1-trifluoroacetone. The PrOF?NMR results reveal roles for the mobile ?3 and L3 regions in the binding of both inhibitors and hydrolyzed ?-lactam products to SPM-1. These results have implications for the mechanisms and inhibition of MBLs by ?-lactams and non-?-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.

SUBMITTER: Abboud MI 

PROVIDER: S-EPMC5396265 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature