Room-temperature structural studies of SARS-CoV-2 protein NendoU with an X-ray free-electron laser.
Room-temperature structural studies of SARS-CoV-2 protein NendoU with an X-ray free-electron laser.
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DOI：
10.1016/j.str.2022.12.009
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发表时间：
2023-02-02
期刊：
影响因子：
5.7
通讯作者：
Fromme, Petra
中科院分区：
文献类型：
--
作者：
Jernigan, Rebecca J.;Logeswaran, Dhenugen;Doppler, Diandra;Nagaratnam, Nirupa;Sonker, Mukul;Yang, Jay-How;Ketawala, Gihan;Martin-Garcia, Jose M.;Shelby, Megan L.;Grant, Thomas D.;Mariani, Valerio;Tolstikova, Alexandra;Sheikh, Michelle Z.;Yung, Mimi Cho;Coleman, Matthew A.;Zaare, Sahba;Kaschner, Emily K.;Rabbani, Mohammad Towshif;Nazari, Reza;Zacks, Michele A.;Hayes, Brandon;Sierra, Raymond G.;Hunter, Mark S.;Lisova, Stella;Batyuk, Alexander;Kupitz, Christopher;Boutet, Sebastien;Hansen, Debra T.;Kirian, Richard A.;Schmidt, Marius;Fromme, Raimund;Frank, Matthias;Ros, Alexandra;Chen, Julian J. -L.;Botha, Sabine;Fromme, Petra
SARS-CoV-2的NendU通过切割反义病毒RNA的多尿苷前导序列来逃避天然免疫系统的能力。在这里，我们报道了NendU的室温结构，用系列飞秒结晶学在2.6欧拉分辨率的X射线自由电子激光器上进行了求解。室温结构提供了对NendU的灵活性、动力学和其他内在属性的洞察，有迹象表明该酶作为变构开关发挥作用。检测溶液和晶体中切割特异性的功能研究支持尿苷-嘌呤切割偏好，我们证明酶活性完全保持在晶体形式。优化了NendU的提纯条件，确定了合适的结晶条件，为未来的时间分辨系列飞秒结晶学研究奠定了基础。这可能会推动设计出治疗冠状病毒感染更有效的抗病毒药物，因为阻止变构构象变化的药物不太容易产生耐药性。SARS-CoV-2的NendU是一种蛋白质，负责病毒逃避免疫系统的能力。Jernigan等人。报告NendU的室温结构，发现有迹象表明它通过交替开关机制发挥作用。这为针对这种酶治疗冠状病毒感染的研究设定了基准。
NendoU from SARS-CoV-2 is responsible for the virus’s ability to evade the innate immune system by cleaving the polyuridine leader sequence of antisense viral RNA. Here we report the room-temperature structure of NendoU, solved by serial femtosecond crystallography at an X-ray free-electron laser to 2.6 Å resolution. The room-temperature structure provides insight into the flexibility, dynamics, and other intrinsic properties of NendoU, with indications that the enzyme functions as an allosteric switch. Functional studies examining cleavage specificity in solution and in crystals support the uridine-purine cleavage preference, and we demonstrate that enzyme activity is fully maintained in crystal form. Optimizing the purification of NendoU and identifying suitable crystallization conditions set the benchmark for future time-resolved serial femtosecond crystallography studies. This could advance the design of antivirals with higher efficacy in treating coronaviral infections, since drugs that block allosteric conformational changes are less prone to drug resistance. NendoU from SARS-CoV-2 is a protein responsible for the virus’s ability to evade the immune system. Jernigan et al. report the room-temperature structure of NendoU, finding indications that it functions by an alternate-switch mechanism. This sets a benchmark for studies targeting this enzyme in treating coronavirus infections.
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发表时间：
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期刊：
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影响因子：
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发表时间：
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影响因子：
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