环索奈德通过靶向HIV-1衣壳蛋白六聚体调节其体外装配  

Ciclesonide acts as a moderator of HIV-1 capsid assembly by targeting the capsid hexamer

作　　者：张莉 张大为 包小峰 ZHANG Li;ZHANG Dawei;BAO Xiaofeng(School of Pharmacy,Nantong University,Nantong 226001,China;Institute of Bioinformatics and Medical Engineering,Jiangsu University of Technology,Changzhou 213001,China)

机构地区：[1]南通大学药学院,南通226001 [2]江苏理工学院生物信息与医药工程研究所,常州213001

出　　处：《生物学杂志》2023年第6期19-24,共6页Journal of Biology

基　　金：国家自然科学基金项目(31700297);南通市第六期江海英才市级培养专项。

摘　　要：多肽CAI在HIV-1衣壳蛋白(CA)C末端的结合口袋可用来筛选CA蛋白装配调节剂。采用均相时间分辨荧光技术(HTRF)筛选399个化合物,发现1个小分子化合物环索奈德(ciclesonide)能够竞争性抑制十二肽CAI与CA蛋白C末端的结合,其半数抑制浓度IC_(50)值为6.06μmol/L。生物膜干涉技术(BLI)测量环索奈德与CA蛋白(单体和六聚体)的结合,环索奈德与CA六聚体的亲和力为159 nmol/L,远远大于其与CA单体的亲和力(K_(D)=1.68 mmol/L)。衣壳蛋白体外装配实验表明,环索奈德能促进CA蛋白的装配,且具有浓度依赖性。分子模拟分析环索奈德与CA蛋白的结合模式,显示环索奈德与CA六聚体的相互作用发生在六聚体相邻亚基N末端-C末端的界面上,并与H67和L211两个残基形成氢键。研究表明,环索奈德是一种潜在的HIV-1衣壳装配调节剂。The CAI-binding pocket in HIV-1 CA CTD can be used as target for antiviral development.In the present study,399 compounds were screened against CA CTD-CAI interaction by a homogeneous time-resolved fluorescence(HTRF)assay.A compound,ciclesonide,was identified as inhibitor by competing with CAI binding to CA CTD(IC_(50)=6.06μmol/L).Biolayer interferometry assay demonstrated that ciclesonide preferentially binded to CA hexamer(K_(D)=159 nmol/L)other than monomer(K_(D)=1.68 mmol/L).In the CA assembly assay,ciclesonide could accelerate the assembly in a concentration-dependent manner.Molecular simulation suggested that ciclesonide could bind to NTD-CTD interface of adjacent protomers in the CA hexamer via H67 and L211 residues.These results indicated that ciclesonide was a potential moderator of HIV-1 capsid assembly.

关 键 词：环索奈德 衣壳装配 C端结构域 人类免疫缺陷病毒1型 均相时间分辨荧光技术 

分 类 号：Q51[生物学—生物化学] R512.91[医药卫生—内科学]