利用荧光分析法测定半合成氨基糖苷类药物与代表细菌和线粒体 A 位的小 RNA 的相对亲和力。
Use of a fluorescence assay to determine relative affinities of semisynthetic aminoglycosides to small RNAs representing bacterial and mitochondrial A sites.
机构信息
Department of Chemistry, Wayne State University, Detroit, MI 48202, USA.
Department of Chemistry, Wayne State University, Detroit, MI 48202, USA.
出版信息
Bioorg Med Chem. 2019 Nov 15;27(22):115121. doi: 10.1016/j.bmc.2019.115121. Epub 2019 Sep 13.
Abstract
The off-target binding of aminoglycosides (AGs) to the A site of human mitochondrial ribosomes in addition to bacterial ribosomes causes ototoxicity and limits their potential as antibiotics. A fluorescence assay was employed to determine relative binding affinities of classical and improved AG compounds to synthetic RNA constructs representing the bacterial and mitochondrial A sites. Results compared well with previously reported in vitro translation assays with engineered ribosomes. Therefore, the minimal RNA motifs and fluorescence assay are shown here to be useful for assessing the selectivity of new compounds.
摘要
除了细菌核糖体，氨基糖苷类（AGs）与人类线粒体核糖体的 A 位结合会导致耳毒性，限制了它们作为抗生素的潜力。荧光测定法被用来确定经典和改良的 AG 化合物与代表细菌和线粒体 A 位的合成 RNA 构建体的相对结合亲和力。结果与先前报道的用工程核糖体进行的体外翻译测定结果吻合较好。因此，这里展示的最小 RNA 基序和荧光测定法可用于评估新化合物的选择性。
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2
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3
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