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| 1 | Nsp2 has the potential to be a drug target revealed by global identification of SARS-CoV-2 Nsp2-interacting proteins显示文摘Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has become a global health threat since December 2019,and there is still no highly effective drug to control the pandemic.To facilitate drug target identification for drug development,studies on molecular mechanisms,such as SARS-CoV-2 protein interactions,are urgently needed.In this study,we focused on Nsp2,a non-structural protein with largely unknown function and mechanism.The interactome of Nsp2 was revealed through the combination of affinity purification mass spectrometry(AP-MS)and stable isotope labeling by amino acids in cell culture(SILAC),and 84 proteins of high-confidence were identified.Gene ontology analysis demonstrated that Nsp2-interacting proteins are involved in several biological processes such as endosome transport and translation.Network analysis generated two clusters,including ribosome assembly and vesicular transport.Bio-layer interferometry(BLI)assay confirmed the bindings between Nsp2-and 4-interacting proteins,i.e.STAU2(Staufen2),HNRNPLL,ATP6V1B2,and RAP1GDS1(SmgGDS),which were randomly selected from the list of 84 proteins.Our findings provide insights into the Nsp2–host interplay and indicate that Nsp2 may play important roles in SARS-CoV-2 infection and serve as a potential drug target for anti-SARS-CoV-2 drug development. | Yun-Xiao Zheng Lei Wang Wei-Sha Kong Hong Chen Xue-Ning Wang Qingfeng Meng Hai-Nan Zhang Shu-Juan Guo He-Wei Jiang Sheng-Ce Tao | 2021 | Acta Biochimica et Biophysica Sinica2021，53，9： | 3 |
| 2 | SARS-CoV-2非结构蛋白Nsp2上调A549细胞泛素水平并被泛素化修饰研究显示文摘目的 新型冠状病毒(SARS-CoV-2)因其强大的感染力肆虐全球,因此深入研究病毒与宿主的相互作用是揭示SARS-CoV-2致病机制的重要途经。方法 利用同源重组方法 将SRAS-CoV-2的非结构蛋白Nsp1~Nsp16构建到真核表达载体上,并通过Western blot检测Nsp1~Nsp16重组质粒的蛋白表达情况。接着使用Western blot检测Nsp2对细胞总泛素水平的影响,并通过细胞免疫荧光检测Nsp2在细胞中的分布情况。最后通过免疫共沉淀技术对Nsp2与泛素之间的相互作用关系进行检测。结果 Western blot检测显示Nsp1~Nsp16重组质粒能够在A549细胞中正常表达;筛选发现非结构蛋白Nsp2能在细胞质中上调细胞总泛素表达水平;免疫共沉淀检测进一步发现Nsp2能与泛素蛋白发生相互作用。结论 成功构建了SARS-CoV-2非结构蛋白Nsp1~Nsp16真核细胞表达载体,并发现非结构蛋白Nsp2可以增加细胞总泛素表达水平并被泛素化修饰。提示Nsp2通过泛素途径参与到SARS-CoV-2与宿主的相互作用,有助于深入了解病毒与宿主的作用机制。 | 游强 张红 宗珊 汪圆松 王承海 李卫玲 孙宾莲 乔嘉璐 | 2023 | 江汉大学学报(自然科学版)2023，51，2： | 0 |
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