Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication.
Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication.
复制标题
氧化应激将 3CLpro 转化为不溶性且更活跃的形式，以促进 SARS-CoV-2 复制
DOI：
10.1016/j.redox.2021.102199
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发表时间：
2021-11-26
期刊：
影响因子：
11.4
通讯作者：
Pan JA
中科院分区：
文献类型：
--
作者：
Du L;Xie Y;Zheng K;Wang N;Gao M;Yu T;Cao L;Shao Q;Zou Y;Xia W;Fang Q;Zhao B;Guo D;Peng X;Pan JA
3CLPro是SARS-CoV-2复制的关键蛋白酶，是抗病毒药物开发的重要靶点。然而，它的活性是如何在细胞内调节的仍不清楚。在这项研究中，我们建立了一个3CLPro蛋白酶活性报告系统，以检测各种因素，包括营养补充剂、离子、PHS或氧化应激诱导剂对3CLPro蛋白酶活性的影响。我们发现，氧化应激可以增加3CLPro的整体活性。在没有改变表达的情况下，氧化应激降低了3CLPro在含有1%Triton-X-100的裂解缓冲液中的溶解度。Triton-X-100不溶的3CLPro与聚集体的形成有关，是酶活性增加的原因。3CLPro蛋白的不溶性和聚集性是由于3CLPro蛋白在Cys85位点之间形成的二硫键所致。除了受到氧化应激的调节外，3CLPro还通过调节GPX1的N端裂解来损害细胞的抗氧化能力。这种切割可以进一步提高3CLPro的氧化活性，有利于3CLPro的聚集和激活，从而导致一个正反馈循环。综上所述，我们报道了氧化应激将3CLPro转化为一种更具酶活性的不溶于洗涤剂的形式，导致病毒复制/转录增加。我们的研究提供了机制证据，表明抗氧化剂在临床治疗新冠肺炎患者中的治疗潜力。
3CLpro is a key proteinase for SARS-CoV-2 replication and serves as an important target for antiviral drug development. However, how its activity is regulated intracellularly is still obscure. In this study, we developed a 3CLpro protease activity reporter system to examine the impact of various factors, including nutrient supplements, ions, pHs, or oxidative stress inducers, on 3CLpro protease activity. We found that oxidative stress could increase the overall activity of 3CLpro. Not altering the expression, oxidative stress decreased the solubility of 3CLpro in the lysis buffer containing 1% Triton-X-100. The Triton-X-100-insoluble 3CLpro was correlated with aggregates’ formation and responsible for the increased enzymatic activity. The disulfide bonds formed between Cys85 sites of 3CLpro protomers account for the insolubility and the aggregation of 3CLpro. Besides being regulated by oxidative stress, 3CLpro impaired the cellular antioxidant capacity by regulating the cleavage of GPx1 at its N-terminus. This cleavage could further elevate the 3CLpro-proximate oxidative activity, favor aggregation and activation of 3CLpro, and thus lead to a positive feedback loop. In summary, we reported that oxidative stress transforms 3CLpro into a detergent-insoluble form that is more enzymatically active, leading to increased viral replication/transcription. Our study provided mechanistic evidence that suggests the therapeutic potential of antioxidants in the clinical treatment of COVID-19 patients.
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影响因子：
16.8
作者：
Jin, Zhenming;Zhao, Yao;Rao, Zihe
通讯作者：
Rao, Zihe
DOI：
10.1038/nrmicro.2016.81
发表时间：
2016-08
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
de Wit E;van Doremalen N;Falzarano D;Munster VJ
通讯作者：
Munster VJ
影响因子：
16
作者：
Pan JA;Sun Y;Jiang YP;Bott AJ;Jaber N;Dou Z;Yang B;Chen JS;Catanzaro JM;Du C;Ding WX;Diaz-Meco MT;Moscat J;Ozato K;Lin RZ;Zong WX
通讯作者：
Zong WX
影响因子：
5.9
作者：
Mody V;Ho J;Wills S;Mawri A;Lawson L;Ebert MCCJC;Fortin GM;Rayalam S;Taval S
通讯作者：
Taval S
影响因子：
5.2
作者：
Alexpandi, Rajaiah;De Mesquita, Joelma Freire;Ravi, Arumugam Veera
通讯作者：
Ravi, Arumugam Veera