New techniques in antibiotic discovery and resistance: Raman spectroscopy
New techniques in antibiotic discovery and resistance: Raman spectroscopy
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DOI：
10.1111/nyas.12847
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发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Heidari-Torkabadi, Hossein
中科院分区：
文献类型：
--
作者：
Carey, Paul R.;Heidari-Torkabadi, Hossein
拉曼光谱在抗生素发现和了解耐药性的分子基础方面都可以发挥作用。药物开发的一个主要挑战是测量细胞系内药物分子的数量，并跟踪它们与细胞内靶点反应的化学反应。最近，一个协议的基础上拉曼显微镜已经开发，实现这些目标。将候选药物浸泡到活细菌细胞中，随后将细胞冷冻并冻干。样品产生示例性（非共振）拉曼数据，其提供每个细胞内药物分子数量的测量以及药物靶点相互作用的细节。结果讨论了两类化合物抑制β-内酰胺酶或二氢叶酸还原酶在一些革兰氏阳性或革兰氏阴性细胞系。本方案的优点是它不使用标记，并且它可以在分钟的时间尺度上测量细胞化合物摄取的动力学。体外拉曼实验支持光谱解释。研究水溶液和单晶中的药物靶标相互作用可以提供对药物靶标相互作用的分子水平的见解，这反过来又为我们理解细菌细胞的数据提供了基础。因此，可以测试X射线晶体学衍生数据对电池内化学的适用性。
Raman spectroscopy can play a role in both antibiotic discovery and understanding the molecular basis of resistance. A major challenge in drug development is to measure the population of the drug molecules inside a cell line and to follow the chemistry of their reactions with intracellular targets. Recently, a protocol based on Raman microscopy has been developed that achieves these goals. Drug candidates are soaked into live bacterial cells and subsequently the cells are frozen and freeze-dried. The samples yield exemplary (nonresonance) Raman data that provide a measure of the number of drug molecules within each cell, as well as details of drug target interactions. Results are discussed for two classes of compounds inhibiting either beta-lactamase or dihydrofolate reductase enzymes in a number of Gram-positive or Gram-negative cell lines. The advantages of the present protocol are that it does not use labels and it can measure the kinetics of cell compound uptake on the time scale of minutes. Spectroscopic interpretation is supported by in vitro Raman experiments. Studying drug target interactions in aqueous solution and in single crystals can provide molecular level insights into drug target interactions, which, in turn, provide the underpinnings of our understanding of data from bacterial cells. Thus, the applicability of X-ray crystallographic derived data to in-cell chemistry can be tested.