通过分裂荧光素酶互补测定鉴定靶向乙型肝炎病毒核心蛋白

StephenW

Antimicrob Agents Chemother. 2018 Sep 17. pii: AAC.01302-18. doi: 10.1128/AAC.01302-18. [Epub ahead of print]
Identification of Compounds Targeting Hepatitis B Virus Core Protein Dimerization Through a Split Luciferase Complementation Assay.
Wei XF1,2, Gan CY1, Cui J1, Luo YY1, Cai XF1, Yuan Y1, Shen J1, Li ZY, Zhang WL1, Long QX1, Hu Y1, Chen J1, Tang N1, Guo H3, Huang AL4,2, Hu JL4,2.
Author information

1
    Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Department of Infectious Diseases, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.
2
    Collaborative Innovation Center for diagnosis and treatment of infectious diseases (CCID), Hangzhou, China.
3
    Department of Microbiology and Immunology, Indiana University School of Medicine.
4
    Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Department of Infectious Diseases, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, China [email protected] [email protected].

Abstract

The capsid of hepatitis B virus is an attractive antiviral target for developing therapies against chronic hepatitis B. Currently available core protein allosteric modulators (CpAMs) mainly affect one of the two major types of protein-protein interactions involved in the process of capsid assembly, which is the interaction between the core dimers. Compounds targeting the interaction between two core monomers have not been rigorously screened due to the lack of screening models. We report herein a cell-based assay in which the formation of core dimers is indicated by the Split Luciferase Complementation (SLC). Making use of this model, 2 compounds, Arbidol and 20-Deoxyingenol, were identified as core dimerization regulators from a library containing 672 compounds. Arbidol and 20-Deoxyingenol inhibit the HBV DNA replication in vitro by decreasing and increasing the formation of core dimer and capsid, respectively. Our results provided a proof of concept for the cell model to be used to screen new agents targeting the step of core dimer and capsid formation.

PMID:
    30224531
DOI:
    10.1128/AAC.01302-18

StephenW

抗微生物剂Chemother。 2018年9月17日.pii：AAC.01302-18。 doi：10.1128 / AAC.01302-18。 [提前打印]
通过分裂荧光素酶互补测定鉴定靶向乙型肝炎病毒核心蛋白质二聚化的化合物。
Wei XF1,2，Gan CY1，Cui J1，Luo YY1，Cai XF1，Yuan Y1，Shen J1，Li ZY，Zhang WL1，Long QX1，Hu Y1，Chen J1，Tang N1，Guo H3，Huang AL4,2，Hu JL4,2。
作者信息

1
    重庆医科大学附属第二医院感染科，传染病分子生物学教育部重点实验室，重庆
2
    中国杭州市传染病诊断与治疗协同创新中心（CCID）。
3
    印第安纳大学医学院微生物学和免疫学系。
4
    重庆医科大学附属第二医院传染病分子生物学教育部重点实验室，重庆，hujieli1977 @ 163.com [email protected]。

抽象

乙型肝炎病毒衣壳是开发针对慢性乙型肝炎的疗法的有吸引力的抗病毒靶标。目前可用的核心蛋白质变构调节剂（CpAM）主要影响衣壳装配过程中涉及的两种主要类型的蛋白质 - 蛋白质相互作用中的一种。是核心二聚体之间的相互作用。由于缺乏筛选模型，未严格筛选靶向两种核心单体之间相互作用的化合物。我们在此报道基于细胞的测定，其中通过分裂荧光素酶互补（SLC）指示核心二聚体的形成。利用该模型，从含有672种化合物的文库中鉴定出2种化合物，即Arbidol和20-Deoxyingenol作为核心二聚化调节剂。 Arbidol和20-Deoxyingenol分别通过减少和增加核心二聚体和衣壳的形成来抑制体外HBV DNA复制。我们的结果提供了细胞模型的概念证明，用于筛选靶向核心二聚体和衣壳形成步骤的新试剂。

结论：
    30224531
DOI：
    10.1128 / AAC.01302-18