Target-based identification of whole-cell active inhibitors of biotin biosynthesis in Mycobacterium tuberculosis.
Target-based identification of whole-cell active inhibitors of biotin biosynthesis in Mycobacterium tuberculosis.
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DOI：
10.1016/j.chembiol.2014.11.012
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发表时间：
2015-01-22
影响因子：
--
通讯作者：
Aldrich CC
中科院分区：
文献类型：
--
作者：
Park SW;Casalena DE;Wilson DJ;Dai R;Nag PP;Liu F;Boyce JP;Bittker JA;Schreiber SL;Finzel BC;Schnappinger D;Aldrich CC
生物素的生物合成是结核分枝杆菌（Mycobacterium tuberculosis，Mtb）在体内生存和存活的关键。氨基转移酶BioA催化生物素途径中的倒数第二步，由于其易受化学抑制，已被确定为有希望的靶点。我们进行了高通量筛选（HTS）采用荧光置换试验，并确定了一套多样性的有效抑制剂，包括许多多样性导向的合成（DOS）支架。为了有效地仅选择靶向生物素生物合成的命中，我们然后在无生物素和含生物素的培养基中针对野生型Mtb并与具有差异水平的BioA表达的同基因BioA Mtb菌株平行地部署全细胞反筛选。事实证明，这种反筛选对于过滤出全细胞活性脱靶的化合物以及鉴定BioA耗尽菌株中具有弱但可测量的全细胞活性的命中至关重要。几个最有希望的命中与BioA共结晶，为未来基于结构的药物设计工作提供框架。
Biotin biosynthesis is essential for survival and persistence of Mycobacterium tuberculosis (Mtb) in vivo. The aminotransferase BioA, which catalyzes the antepenultimate step in the biotin pathway, has been established as a promising target due to its vulnerability to chemical inhibition. We performed high-throughput screening (HTS) employing a fluorescence displacement assay and identified a diverse set of potent inhibitors including many diversity-oriented synthesis (DOS) scaffolds. To efficiently select only hits targeting biotin biosynthesis, we then deployed a whole-cell counter-screen in either biotin-free and biotin-containing medium against wild-type Mtb and in parallel with isogenic bioA Mtb strains that possess differential levels of BioA expression. This counter-screen proved crucial to filter out compounds whose whole-cell activity was off-target as well as identify hits with weak, but measurable whole-cell activity in BioA-depleted strains. Several of the most promising hits were co-crystallized with BioA to provide a framework for future structure-based drug design efforts.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
DOI：
10.1021/jo4000916
发表时间：
2013-06-07
期刊：
The Journal of organic chemistry
影响因子：
--
作者：
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通讯作者：
Marcaurelle LA