Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Methods Enzymol. 2022;663:259-271. doi: 10.1016/bs.mie.2021.09.008. Epub 2021 Dec 28.
Antimicrobial resistance (AMR) is a significant public health issue that threatens our ability to treat common infections. AMR often emerges in bacteria through upregulation of proteins that allow a subpopulation of resistant bacteria to proliferate through natural selection. Identifying these proteins is crucial for understanding how AMR develops in bacteria and is essential in developing novel therapeutics to combat the threat of widespread AMR. Mass spectrometry-based proteomics is a powerful tool for understanding the biochemical pathways of biological systems, lending remarkable insight into AMR mechanisms in bacteria through measuring the changing protein abundances as a result of antibiotic treatment. Here, we describe a serial passaging method for evolving resistance in bacteria that implements quantitative proteomics to reveal the differential proteomes of resistant bacteria. The focus herein is on antimicrobial peptides (AMPs), but the approach can be generalized for any antimicrobial compound. Comparative proteomics of sensitive vs. resistance strains in response to AMP treatment reveals mechanisms to survive the bioactive compound and points to the mechanism of action for novel AMPs.
抗微生物药物耐药性（AMR）是一个重大的公共卫生问题，威胁着我们治疗常见感染的能力。AMR 通常通过上调允许一小部分耐药细菌通过自然选择增殖的蛋白质在细菌中出现。鉴定这些蛋白质对于了解细菌中 AMR 的发展至关重要，并且对于开发新型治疗方法来对抗广泛存在的 AMR 威胁至关重要。基于质谱的蛋白质组学是理解生物系统生化途径的强大工具，通过测量抗生素治疗后蛋白质丰度的变化，为细菌中的 AMR 机制提供了显著的见解。在这里，我们描述了一种在细菌中进行耐药性进化的连续传代方法，该方法采用定量蛋白质组学来揭示耐药细菌的差异蛋白质组。本文的重点是抗菌肽（AMP），但该方法可以推广到任何抗菌化合物。对 AMP 处理后敏感和耐药菌株的比较蛋白质组学揭示了存活于生物活性化合物的机制，并指出了新型 AMP 的作用机制。
