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胞嘧啶肽霉素与烟草花叶病毒超家族1解旋酶蛋白之间的结合研究。

Binding studies between cytosinpeptidemycin and the superfamily 1 helicase protein of tobacco mosaic virus.

作者信息

Li Xiangyang, Chen Kai, Gao Di, Wang Dongmei, Huang Maoxi, Zhu Hengmin, Kang Jinxin

机构信息

State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University Guiyang 550025 PR China

出版信息

RSC Adv. 2018 May 23;8(34):18952-18958. doi: 10.1039/c8ra01466c. eCollection 2018 May 22.

DOI:10.1039/c8ra01466c
PMID:35539684
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9080650/
Abstract

Tobacco mosaic virus (TMV) helicases play important roles in viral multiplication and interactions with host organisms. They can also be targeted by antiviral agents. Cytosinpeptidemycin has a good control effect against TMV. However, the mechanism of action is unclear. In this study, we expressed and purified TMV superfamily 1 helicase (TMV-Hel) and analyzed its three-dimensional structure. Furthermore, the binding interactions of TMV-Hel and cytosinpeptidemycin were studied. Microscale thermophoresis and isothermal titration calorimetry experiments showed that cytosinpeptidemycin bound to TMV-Hel with a dissociation constant of 0.24-0.44 μM. Docking studies provided further insights into the interaction of cytosinpeptidemycin with the His375 of TMV-Hel. Mutational and Microscale thermophoresis analyses showed that cytosinpeptidemycin bound to a TMV-Hel mutant (H375A) with a dissociation constant of 14.5 μM. Thus, His375 may be the important binding site for cytosinpeptidemycin. The data are important for designing and synthesizing new effective antiphytoviral agents.

摘要

烟草花叶病毒（TMV）解旋酶在病毒增殖以及与宿主生物体的相互作用中发挥着重要作用。它们也可成为抗病毒药物的作用靶点。胞嘧啶肽霉素对TMV具有良好的防治效果。然而，其作用机制尚不清楚。在本研究中，我们表达并纯化了TMV超家族1解旋酶（TMV-Hel），并分析了其三维结构。此外，还研究了TMV-Hel与胞嘧啶肽霉素的结合相互作用。微量热泳动和等温滴定量热法实验表明，胞嘧啶肽霉素与TMV-Hel结合，解离常数为0.24 - 0.44 μM。对接研究进一步揭示了胞嘧啶肽霉素与TMV-Hel的His375之间的相互作用。突变和微量热泳动分析表明，胞嘧啶肽霉素与TMV-Hel突变体（H375A）结合，解离常数为14.5 μM。因此，His375可能是胞嘧啶肽霉素的重要结合位点。这些数据对于设计和合成新型有效的抗植物病毒药物具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/e3ce2f01cfff/c8ra01466c-f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/84b3383b24b3/c8ra01466c-f1.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/b241a4111da5/c8ra01466c-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/6e2bc9cdd405/c8ra01466c-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/4d77001b215c/c8ra01466c-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/e3ce2f01cfff/c8ra01466c-f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/84b3383b24b3/c8ra01466c-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/4dee4c636005/c8ra01466c-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/2f23e5ced807/c8ra01466c-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/b241a4111da5/c8ra01466c-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/6e2bc9cdd405/c8ra01466c-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/4d77001b215c/c8ra01466c-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30b/9080650/e3ce2f01cfff/c8ra01466c-f7.jpg

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