Crystal structure of the Ilheus virus helicase: implications for enzyme function and drug design

作者全名："Wang, De-Ping; Wang, Mei-Yue; Li, Yong-Mei; Shu, Wen; Cui, Wen; Jiang, Fang-Ying; Zhou, Xin; Wang, Wen-Ming; Cao, Ji-Min"

作者地址："[Wang, De-Ping; Wang, Mei-Yue; Li, Yong-Mei; Shu, Wen; Jiang, Fang-Ying; Zhou, Xin; Cao, Ji-Min] Shanxi Med Univ, Key Lab Cellular Physiol, Minist Educ, Taiyuan, Peoples R China; [Wang, De-Ping; Wang, Mei-Yue; Li, Yong-Mei; Shu, Wen; Jiang, Fang-Ying; Zhou, Xin; Cao, Ji-Min] Shanxi Med Univ, Dept Physiol, Taiyuan, Peoples R China; [Cui, Wen] Chongqing Med Univ, Inst Life Sci, Chongqing, Peoples R China; [Wang, Wen-Ming] Shanxi Univ, Inst Mol Sci, Taiyuan, Peoples R China"

通信作者："Zhou, X; Cao, JM (通讯作者)，Shanxi Med Univ, Key Lab Cellular Physiol, Minist Educ, Taiyuan, Peoples R China.; Zhou, X; Cao, JM (通讯作者)，Shanxi Med Univ, Dept Physiol, Taiyuan, Peoples R China.; Wang, WM (通讯作者)，Shanxi Univ, Inst Mol Sci, Taiyuan, Peoples R China."

来源：CELL AND BIOSCIENCE

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:000782760100001

JCR分区：Q1

影响因子：7.5

年份：2022

卷号：12

期号：1

开始页：　

结束页：　

文献类型：Article

关键词：Ilheus virus; NS3 helicase; Crystal structure; ATP hydrolysis; Molecular docking

摘要："Background The Ilheus virus (ILHV) is an encephalitis associated arthropod-borne flavivirus. It was first identified in Ilheus City in the northeast Brazil before spreading to a wider geographic range. No specific vaccines or drugs are currently available for the treatment of ILHV infections. The ILHV helicase, like other flavivirus helicases, possesses 5MODIFIER LETTER PRIME-triphosphatase activity. This allows it to perform ATP hydrolysis to generate energy as well as sustain double-stranded RNA's unwinding during ILHV genome replication. Thus, ILHV helicase is an ideal target for inhibitor design. Results We determined the crystal structure of the ILHV helicase at 1.75-angstrom resolution. We then conducted molecular docking of ATP-Mn2+ to the ILHV helicase. Comparisons with related flavivirus helicases indicated that both the NTP and the RNA-ILHV helicase binding sites were conserved across intra-genus species. This suggested that ILHV helicase adopts an identical mode in recognizing ATP/Mn2+. However, the P-loop in the active site showed a distinctive conformation; reflecting a different local structural rearrangement. ILHV helicase enzymatic activity was also characterized. This was found to be relatively lower than that of the DENV, ZIKV, MVE, and ALSV helicases. Our structure-guided mutagenesis revealed that R26A, E110A, and Q280A greatly reduced the ATPase activities. Moreover, we docked two small molecule inhibitors of DENV helicase (ST-610 and suramin) to the ILHV helicase and found that these two molecules had the potential to inhibit the activity of ILHV helicase as well. Conclusion High-resolution ILHV helicase structural analysis demonstrates the key amino acids of ATPase activities and could be useful for the design of inhibitors targeting the helicase of ILHV."

基金机构："Key Medical Science and Technology Program of Shanxi Province [2020XM01]; Shanxi ""1331 Project"" Quality and Efficiency Improvement Plan [1331KFC]; National Natural Science Foundation of China [82170523, 81801858]"

基金资助正文："This study was supported by Key Medical Science and Technology Program of Shanxi Province (2020XM01), Shanxi ""1331 Project"" Quality and Efficiency Improvement Plan (1331KFC), and National Natural Science Foundation of China (82170523, 81801858)."