19 F?NMR Monitoring of Reversible Protein Post-Translational Modifications: Class?D ?-Lactamase Carbamylation and Inhibition.

ABSTRACT: Bacterial production of ?-lactamases with carbapenemase activity is a global health threat. The active sites of class?D carbapenemases such as OXA-48, which is of major clinical importance, uniquely contain a carbamylated lysine residue which is essential for catalysis. Although there is significant interest in characterizing this post-translational modification, and it is a promising inhibition target, protein carbamylation is challenging to monitor in solution. We report the use of 19 F?NMR spectroscopy to monitor the carbamylation state of 19 F-labelled OXA-48. This method was used to investigate the interactions of OXA-48 with clinically used serine ?-lactamase inhibitors, including avibactam and vaborbactam. Crystallographic studies on 19 F-labelled OXA-48 provide a structural rationale for the sensitivity of the 19 F label to active site interactions. The overall results demonstrate the use of 19 F?NMR to monitor reversible covalent post-translational modifications.

SUBMITTER: van Groesen E 

PROVIDER: S-EPMC6771976 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature