Multiple unbiased approaches identify oxidosqualene cyclase as the molecular target of a promising anti-leishmanial.
Multiple unbiased approaches identify oxidosqualene cyclase as the molecular target of a promising anti-leishmanial.
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DOI：
10.1016/j.chembiol.2021.02.008
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发表时间：
2021-05-20
影响因子：
8.6
通讯作者：
Wyllie S
中科院分区：
文献类型：
--
作者：
Paradela LS;Wall RJ;Carvalho S;Chemi G;Corpas-Lopez V;Moynihan E;Bello D;Patterson S;Güther MLS;Fairlamb AH;Ferguson MAJ;Zuccotto F;Martin J;Gilbert IH;Wyllie S
表型筛选鉴定出一种对内脏利什曼病病原体多诺瓦利什曼原虫有活性的苯并噻吩化合物。采用多重正交法，确定了甾醇生物合成的关键酶氧化角鲨烯环化酶（OSC）为该外消旋化合物及其对映体的靶酶。全基因组测序和全基因组过表达文库筛选证实，OSC基因扩增与对化合物1的抗性有关。在野生型细胞中引入OSC基因的异位拷贝降低了对这些化合物的易感性，证实了该酶在抗性中的作用。生化分析表明，化合物(S)-1处理的promastigotes和无细胞膜制备中，OSC底物的积累和产物的消耗分别发生在化合物(S)-1处理的promastigotes中。热蛋白质组分析证实化合物(S)-1直接与OSC结合。最后，建模和对接研究确定了化合物(S)-1与LdOSC活性位点之间的关键相互作用。提出了提高这种有前途的抗利什曼原虫效力的策略。遗传学和化学蛋白质组学确定了一种有希望的抗利什曼原虫的靶标，生化分析证实了细胞中氧化角鲨烯环化酶的直接抑制作用，对接和建模研究确定了化合物与靶标之间的关键相互作用，提出了提高这种苯并噻吩效力的策略，Paradela等人研究了一种有希望的抗利什曼原虫的作用机制。利用遗传学、化学蛋白质组学和生化等方法，证实该苯并噻吩的作用靶点为氧化角鲨烯环化酶，是甾醇生物合成的关键酶。对接和建模确定了该化合物与该酶活性位点之间的关键相互作用。
Phenotypic screening identified a benzothiophene compound with activity against Leishmania donovani, the causative agent of visceral leishmaniasis. Using multiple orthogonal approaches, oxidosqualene cyclase (OSC), a key enzyme of sterol biosynthesis, was identified as the target of this racemic compound and its enantiomers. Whole genome sequencing and screening of a genome-wide overexpression library confirmed that OSC gene amplification is associated with resistance to compound 1. Introduction of an ectopic copy of the OSC gene into wild-type cells reduced susceptibility to these compounds confirming the role of this enzyme in resistance. Biochemical analyses demonstrated the accumulation of the substrate of OSC and depletion of its product in compound (S)-1-treated-promastigotes and cell-free membrane preparations, respectively. Thermal proteome profiling confirmed that compound (S)-1 binds directly to OSC. Finally, modeling and docking studies identified key interactions between compound (S)-1 and the LdOSC active site. Strategies to improve the potency for this promising anti-leishmanial are proposed. Genetics and chemo-proteomics identify the target of a promising anti-leishmanial Biochemical assays confirm the direct inhibition of oxidosqualene cyclase in cells Docking and modeling studies identify key interactions between compound and target Strategies to improve the potency of this benzothiophene are proposed Paradela et al. investigated the mechanism of action of a promising anti-leishmanial. Using genetics, chemo-proteomics, and biochemical approaches, the target of this benzothiophene was confirmed as oxidosqualene cyclase, a key enzyme of sterol biosynthesis. Docking and modeling identified key interactions between this compound and the active site of this enzyme.
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DOI：
10.4269/ajtmh.1981.30.566
发表时间：
1981-01-01
影响因子：
3.3
作者：
BERMAN, JD
通讯作者：
BERMAN, JD
影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
3.8
作者：
Liang, Yayun;Besch-Williford, Cynthia;Hyder, Salman M.
通讯作者：
Hyder, Salman M.
影响因子：
2.4
作者：
Don, Rob;Ioset, Jean-Robert
通讯作者：
Ioset, Jean-Robert
影响因子：
3.6
作者：
Bahamdan, KA;Tallab, TM;Nauri, MM
通讯作者：
Nauri, MM