Complex Structure of Pseudomonas aeruginosa Arginine Rhamnosyltransferase EarP with Its Acceptor Elongation Factor P
Complex Structure of Pseudomonas aeruginosa Arginine Rhamnosyltransferase EarP with Its Acceptor Elongation Factor P
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DOI：
10.1128/jb.00128-19
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发表时间：
2019-07-01
影响因子：
3.2
通讯作者：
Xiao, Yazhong
中科院分区：
文献类型：
--
作者：
He, Chao;Liu, Ning;Xiao, Yazhong
细菌转化糖基转移酶EarP将dTDP-β-L-鼠李糖（TDP-Rha）中的鼠李糖转移到翻译延伸因子P（EF-P）的Arg 32上以激活其功能。我们在这里报告的结构和生化特性的铜绿假单胞菌EarP。与最近报道的脑膜炎奈瑟菌EarP相反，铜绿假单胞菌EarP在TDP-Rha和EF-P结合后表现出不同的构象变化。糖供体结合增强受体与EarP的结合，如通过apo-、TDP-Rha-和TDP/EF-P结合形式之间的结构比较和等温滴定量热实验所揭示的。体外EF-P鼠李糖基化结合活性中心几何构型表明，Asp 16对应于N.脑膜炎EarP是催化碱基，而Glu 272是另一个推定的催化残基。我们的研究为EarP靶向抑制剂的设计提供了基础，该抑制剂可用于对抗铜绿假单胞菌和其他临床相关物种的感染。重要意义EF-P的翻译后鼠李糖基化在铜绿假单胞菌中起着关键作用，建立了毒力和抗生素耐药性，以及生存。来自铜绿假单胞菌的EF-P特异性鼠李糖基转移酶EarP的详细结构和生物化学表征不仅表明糖供体TDP-Rha结合增强受体EF-P与EarP的结合，而且还应为针对铜绿假单胞菌和其他含EarP的病原体的感染的其抑制剂的结构导向开发提供有价值的信息。
A bacterial inverting glycosyltransferase EarP transfers rhamnose from dTDP-beta-L-rhamnose (TDP-Rha) to Arg32 of translation elongation factor P (EF-P) to activate its function. We report here the structural and biochemical characterization of Pseudomonas aeruginosa EarP. In contrast to recently reported Neisseria meningitidis EarP, P. aeruginosa EarP exhibits differential conformational changes upon TDP-Rha and EF-P binding. Sugar donor binding enhances acceptor binding to EarP, as revealed by structural comparison between the apo-, TDP-Rha-, and TDP/EF-P-bound forms and isothermal titration calorimetry experiments. In vitro EF-P rhamnosylation combined with active-site geometry indicates that Asp16 corresponding to Asp20 of N. meningitidis EarP is the catalytic base, whereas Glu272 is another putative catalytic residue. Our study should provide the basis for EarP-targeted inhibitor design against infections from P. aeruginosa and other clinically relevant species.IMPORTANCE Posttranslational rhamnosylation of EF-P plays a key role in Pseudomonas aeruginosa, establishing virulence and antibiotic resistance, as well as survival. The detailed structural and biochemical characterization of the EF-P-specific rhamnosyltransferase EarP from P. aeruginosa not only demonstrates that sugar donor TDP-Rha binding enhances acceptor EF-P binding to EarP but also should provide valuable information for the structure-guided development of its inhibitors against infections from P. aeruginosa and other EarP-containing pathogens.