Aminoglycoside ribosome interactions reveal novel conformational states at ambient temperature.
Aminoglycoside ribosome interactions reveal novel conformational states at ambient temperature.
复制标题
氨基糖苷核糖体相互作用揭示了环境温度下的新构象状态。
DOI：
10.1093/nar/gky693
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发表时间：
2018
影响因子：
14.9
通讯作者：
O
中科院分区：
文献类型：
--
作者：
O'Sullivan,MaryE;Poitevin,Frédéric;Sierra,RaymondG;Gati,Cornelius;Dao,EHan;Rao,Yashas;Aksit,Fulya;Ciftci,Halilibrahim;Corsepius,Nicholas;Greenhouse,Robert;Hayes,Brandon;Hunter,MarkS;Liang,Mengling;McGurk,Alex;Mbgam,Paul;O
细菌 30S 核糖体亚基是主要的抗生素靶点。尽管已有数十年的发现，但抗生素结合引起核糖体功能障碍的机制尚未完全了解。环境温度晶体学技术可以对局部抗生素结合位点相互作用如何触发扰乱蛋白质合成的全局亚基重排进行更生物学相关的研究。在此，研究了 2-脱氧链霉胺（巴龙霉素和西索米星）、一种新型西索米星衍生物、N1-甲基磺酰西索米星 (N1MS) 和非脱氧链霉胺（链霉素）氨基糖苷类在环境温度和低温下对核糖体的结构影响。比较研究得出三个主要观察结果。首先，解码中心中的各个氨基糖苷-核糖体相互作用对于低温结构与环境温度结构是相似的。其次，对 h45 高度保守的 GGAA 四环的分析揭示了氨基糖苷类特异性构象变化，这些变化仅对 N1MS 受温度影响。我们报告了不同状态下的 h44-h45 界面，即接合、脱离和平衡。第三，我们观察了氨基糖苷类药物对 30S 结构域闭合的影响，包括一种新的中间闭合状态，该状态对温度也敏感。对三个常温晶体学和五个低温晶体学数据集的分析揭示了 h44-h45 接合和域闭合之间的相关性。这些观察结果说明了环境温度晶体学在识别由局部药物结合位点相互作用引起的核糖体功能障碍的动态机制中的作用。这些数据共同确定了氨基糖苷结合诱导的三级核糖体结构变化，为药物设计提供了功能见解和目标。
The bacterial 30S ribosomal subunit is a primary antibiotic target. Despite decades of discovery, the mechanisms by which antibiotic binding induces ribosomal dysfunction are not fully understood. Ambient temperature crystallographic techniques allow more biologically relevant investigation of how local antibiotic binding site interactions trigger global subunit rearrangements that perturb protein synthesis. Here, the structural effects of 2-deoxystreptamine (paromomycin and sisomicin), a novel sisomicin derivative, N1-methyl sulfonyl sisomicin (N1MS) and the non-deoxystreptamine (streptomycin) aminoglycosides on the ribosome at ambient and cryogenic temperatures were examined. Comparative studies led to three main observations. First, individual aminoglycoside–ribosome interactions in the decoding center were similar for cryogenic versus ambient temperature structures. Second, analysis of a highly conserved GGAA tetraloop of h45 revealed aminoglycoside-specific conformational changes, which are affected by temperature only for N1MS. We report the h44–h45 interface in varying states, i.e. engaged, disengaged and in equilibrium. Third, we observe aminoglycoside-induced effects on 30S domain closure, including a novel intermediary closure state, which is also sensitive to temperature. Analysis of three ambient and five cryogenic crystallography datasets reveal a correlation between h44–h45 engagement and domain closure. These observations illustrate the role of ambient temperature crystallography in identifying dynamic mechanisms of ribosomal dysfunction induced by local drug-binding site interactions. Together, these data identify tertiary ribosomal structural changes induced by aminoglycoside binding that provides functional insight and targets for drug design.