Department of Chemistry, University of Oxford, Oxford, OX1 3TA, UK.
Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON, K7L 3N6, Canada.
Chemistry. 2019 Sep 12;25(51):11837-11841. doi: 10.1002/chem.201902529. Epub 2019 Aug 20.
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 F NMR spectroscopy to monitor the carbamylation state of F-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 F-labelled OXA-48 provide a structural rationale for the sensitivity of the F label to active site interactions. The overall results demonstrate the use of F NMR to monitor reversible covalent post-translational modifications.
细菌产生具有碳青霉烯酶活性的β-内酰胺酶是全球健康威胁。OXA-48 等 D 类碳青霉烯酶的活性部位独特地含有一个碳化赖氨酸残基，这对于催化是必不可少的。尽管人们对描述这种翻译后修饰很感兴趣，并且它是一个有前途的抑制靶标，但在溶液中监测蛋白质碳化是具有挑战性的。我们报告了使用 F NMR 光谱法监测 F 标记的 OXA-48 的碳化状态。该方法用于研究 OXA-48 与临床使用的丝氨酸β-内酰胺酶抑制剂（包括阿维巴坦和沃博巴坦）的相互作用。F 标记的 OXA-48 的晶体学研究为 F 标记对活性部位相互作用的敏感性提供了结构依据。总体结果表明，使用 F NMR 来监测可逆的共价翻译后修饰。
