Comparative In Vitro and In Silico Analysis of the Selectivity of Indirubin as a Human Ah Receptor Agonist.
Comparative In Vitro and In Silico Analysis of the Selectivity of Indirubin as a Human Ah Receptor Agonist.
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比较体外和硅氧素作为人类AH受体激动剂的选择性的计算机分析。
DOI：
10.3390/ijms19092692
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发表时间：
2018-09-10
影响因子：
5.6
通讯作者：
Denison MS
中科院分区：
文献类型：
--
作者：
Faber SC;Soshilov AA;Giani Tagliabue S;Bonati L;Denison MS
芳烃受体（AhR）是一种配体依赖性转录因子，在其被结构多样的化学物质结合和激活后调节基因表达。已经观察到AhR功能的物种差异，小鼠AhR（mAhR）和人AhR（hAhR）在配体结合、辅激活因子募集、基因表达和应答方面表现出显著差异。虽然AhR激动剂靛玉红（IR）是比原型配体2，3，7，8-四氯二苯并对二恶英（TCDD）更有效的hAhR依赖性基因表达激活剂，但它对mAhR的激活作用明显较弱。DNA结合分析证实了IR在体外刺激hAhR的转化/DNA结合方面的更大效力/功效，并且结构域交换实验表明，对IR的增强响应主要是由于hAhR配体结合结构域（LBD）。定点突变和功能分析研究表明，H326和A349在mAhR LBD中的相应残基的hAhR LBD中的突变显着增加IR的效力。由于这些突变对配体结合没有显着的影响，这些残基可能有助于提高转化/DNA结合的IR结合的hAhR的效率。分子对接mAhR LBD同源性模型进一步阐明了不同的作用A375 V突变TCDD和IR结合，所揭示的[3 H]TCDD竞争性结合的结果。这些结果证明了给定AhR的LBD内结构多样的配体的差异结合，并证实AhR的LBD内的氨基酸差异导致配体反应的显著物种差异。
The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that modulates gene expression following its binding and activation by structurally diverse chemicals. Species differences in AhR functionality have been observed, with the mouse AhR (mAhR) and human AhR (hAhR) exhibiting significant differences in ligand binding, coactivator recruitment, gene expression and response. While the AhR agonist indirubin (IR) is a more potent activator of hAhR-dependent gene expression than the prototypical ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), it is a significantly less potent activator of the mAhR. DNA binding analysis confirmed the greater potency/efficacy of IR in stimulating transformation/DNA binding of the hAhR in vitro and domain-swapping experiments demonstrated that the enhanced response to IR was primarily due to the hAhR ligand binding domain (LBD). Site-directed mutagenesis and functional analysis studies revealed that mutation of H326 and A349 in the mAhR LBD to the corresponding residues in the hAhR LBD significantly increased the potency of IR. Since these mutations had no significant effect on ligand binding, these residues likely contribute to an enhanced efficiency of transformation/DNA binding by IR-bound hAhR. Molecular docking to mAhR LBD homology models further elucidated the different roles of the A375V mutation in TCDD and IR binding, as revealed by [3H]TCDD competitive binding results. These results demonstrate the differential binding of structurally diverse ligands within the LBD of a given AhR and confirm that amino acid differences within the LBD of AhRs contribute to significant species differences in ligand response.
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DOI：
10.1093/toxsci/kfn149
发表时间：
2008-11
期刊：
Toxicological sciences : an official journal of the Society of Toxicology
影响因子：
--
作者：
Bunger MK;Glover E;Moran SM;Walisser JA;Lahvis GP;Hsu EL;Bradfield CA
通讯作者：
Bradfield CA
影响因子：
3.6
作者：
Flaveny, Colin A.;Murray, Iain A.;Perdew, Gary H.
通讯作者：
Perdew, Gary H.
影响因子：
4.6
作者：
Bonati, Laura;Corrada, Dario;Motta, Stefano
通讯作者：
Motta, Stefano
影响因子：
6.1
作者：
Baston, David S.;Denison, Michael S.
通讯作者：
Denison, Michael S.
影响因子：
3.8
作者：
Adachi, J;Mori, Y;Matsuda, T
通讯作者：
Matsuda, T