Allosteric regulation and crystallographic fragment screening of SARS-CoV-2 NSP15 endoribonuclease.
Allosteric regulation and crystallographic fragment screening of SARS-CoV-2 NSP15 endoribonuclease.
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DOI：
10.1093/nar/gkad314
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发表时间：
2023-06-09
影响因子：
14.9
通讯作者：
Oliva, Glaucius
中科院分区：
文献类型：
--
作者：
Godoy, Andre Schutzer;Nakamura, Aline Minalli;Douangamath, Alice;Song, Yun;Noske, Gabriela Dias;Gawriljuk, Victor Oliveira;Fernandes, Rafaela Sachetto;Pereira, Humberto D. Muniz;Oliveira, Ketllyn Irene Zagato;Fearon, Daren;Dias, Alexandre;Krojer, Tobias;Fairhead, Michael;Powell, Alisa;Dunnet, Louise;Brandao-Neto, Jose;Skyner, Rachael;Chalk, Rod;Bajusz, David;Bege, Miklos;Borbas, Aniko;Keseru, Gyoergy Miklos;von Delft, Frank;Oliva, Glaucius
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是2019冠状病毒病（COVID-19）的病原体。NSP 15内切核糖核酸酶，称为NendoU，是高度保守的，在病毒逃避免疫系统的能力中起着关键作用。NendoU是开发新的抗病毒药物的一个有前途的靶点。然而，酶的结构和动力学的复杂性，沿着广泛的识别序列和缺乏结构复合物，阻碍了抑制剂的开发。在这里，我们进行了NendoU在其单体和六聚体形式的酶特性，显示六聚体是变构酶与积极的合作指数，并与锰的酶活性没有影响。通过结合不同pH下的冷冻电子显微镜、X射线晶体学以及生化和结构分析，我们发现NendoU可以在开放和封闭形式之间转换，这可能分别对应于活性和非活性状态。我们还探讨了NendoU组装成更大的超分子结构的可能性，并提出了变构调节的机制。此外，我们对NendoU进行了大片段筛选活动，并确定了几个新的变构位点，这些位点可以用于开发新的抑制剂。总的来说，我们的研究结果为NendoU的复杂结构和功能提供了见解，并为抑制剂的开发提供了新的机会。SARS-CoV-2 NendoU在活性和非活性构象之间转换，片段筛选显示多个变构位点。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). The NSP15 endoribonuclease enzyme, known as NendoU, is highly conserved and plays a critical role in the ability of the virus to evade the immune system. NendoU is a promising target for the development of new antiviral drugs. However, the complexity of the enzyme's structure and kinetics, along with the broad range of recognition sequences and lack of structural complexes, hampers the development of inhibitors. Here, we performed enzymatic characterization of NendoU in its monomeric and hexameric form, showing that hexamers are allosteric enzymes with a positive cooperative index, and with no influence of manganese on enzymatic activity. Through combining cryo-electron microscopy at different pHs, X-ray crystallography and biochemical and structural analysis, we showed that NendoU can shift between open and closed forms, which probably correspond to active and inactive states, respectively. We also explored the possibility of NendoU assembling into larger supramolecular structures and proposed a mechanism for allosteric regulation. In addition, we conducted a large fragment screening campaign against NendoU and identified several new allosteric sites that could be targeted for the development of new inhibitors. Overall, our findings provide insights into the complex structure and function of NendoU and offer new opportunities for the development of inhibitors. SARS-CoV-2 NendoU shifts between active and inactive conformation, and fragment screening revealed multiple allosteric sites.
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一种具有新故事的“旧”蛋白质：冠状病毒内切核糖核酸酶对于逃避宿主抗病毒防御非常重要。
影响因子：
3.7
作者：
Deng X;Baker SC
通讯作者：
Baker SC
影响因子：
6.4
作者：
Athmer J;Fehr AR;Grunewald M;Smith EC;Denison MR;Perlman S
通讯作者：
Perlman S
影响因子：
16.6
作者：
Douangamath A;Fearon D;Gehrtz P;Krojer T;Lukacik P;Owen CD;Resnick E;Strain-Damerell C;Aimon A;Ábrányi-Balogh P;Brandão-Neto J;Carbery A;Davison G;Dias A;Downes TD;Dunnett L;Fairhead M;Firth JD;Jones SP;Keeley A;Keserü GM;Klein HF;Martin MP;Noble MEM;O'Brien P;Powell A;Reddi RN;Skyner R;Snee M;Waring MJ;Wild C;London N;von Delft F;Walsh MA
通讯作者：
Walsh MA
影响因子：
14.9
作者：
ASLANIDIS, C;DEJONG, PJ
通讯作者：
DEJONG, PJ
DOI：
10.1107/s205979831700331x
发表时间：
2017-03-01
影响因子：
2.2
作者：
Collins, Patrick M.;Ng, Jia Tsing;von Delft, Frank
通讯作者：
von Delft, Frank