19F‐NMR monitoring of reversible protein post‐translational modifications: Class D β‐lactamase carbamylation and inhibition

DOI: 10.1002/chem.201902529

Authors: Emma Van Groesen (University of Oxford) , Christopher T. Lohans (University of Oxford) , Jurgen Brem (University of Oxford) , Kristina M. J. Aertker (University of Oxford) , Timothy D. W. Claridge (University of Oxford) , Christopher Schofield (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Chemistry – A European Journal

State: Published (Approved)
Published: July 2019
Diamond Proposal Number(s): 12346

Abstract: Bacterial production of β‐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 19F‐NMR spectroscopy to monitor the carbamylation state of 19F‐labelled OXA‐48. This method was used to investigate the interactions of OXA‐48 with clinically used serine β‐ lactamase inhibitors, including avibactam and vaborbactam. Crystallographic studies on 19F‐labelled OXA‐48 provide a structural rationale for the sensitivity of the 19F‐label to active site interactions. The overall results demonstrate the use of 19F‐NMR to monitor reversible covalent post‐translational modifications.

Journal Keywords: antibiotics; carbamylation; carbapenemase; β-lactamase; NMR spectroscopy

Diamond Keywords: Bacteria

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)

Added On: 31/07/2019 11:37

Documents:
chem.201902529.pdf

Discipline Tags:

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

Technical Tags:

Diffraction Macromolecular Crystallography (MX)