The binding sites for benztropines and dopamine in the dopamine transporter overlap.
The binding sites for benztropines and dopamine in the dopamine transporter overlap.
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DOI：
10.1016/j.neuropharm.2010.08.021
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发表时间：
2011-01
影响因子：
4.7
通讯作者：
Gether, Ulrik
中科院分区：
文献类型：
--
作者：
Bisgaard, Heidi;Larsen, M. Andreas B.;Mazier, Sonia;Beuming, Thijs;Newman, Amy Hauck;Weinstein, Hare;Shi, Lei;Loland, Claus J.;Gether, Ulrik
关键词：
苯托品（BZTs）的类似物是多巴胺转运蛋白（DAT）的有效抑制剂，但作为行为兴奋剂不如可卡因有效。因此，人们一直在努力评估这些化合物作为可卡因成瘾潜在药物的先导。在这里，我们使用计算建模与定点诱变的特征BZT在DAT的结合位点。对接到分子模型的基础上的细菌同系物LeuT的结构支持BZT结合位点，与底物结合口袋重叠。一致的是，口袋内的残基突变，包括Val 1523.46 * 突变为Ala或Ile，Ser 4228.60突变为Ala和Asn 1573.51突变为Cys或Ala，导致对BZT和类似物JHW 007的亲和力降低，如在[3 H]多巴胺摄取抑制试验和/或[3 H]CFT竞争结合试验中所评估的。JHW 007中的一个苯环氟取代基与Asn 1573.51的假定极性相互作用被用作确定可能结合位姿的标准，并为诱变结果建立结构背景。分析将JHW 007的另一个氟取代苯基环定位在跨膜片段（TM）10中的Ala47910.51/Ala48010.52附近。与JHW 007相比，BZT缺乏这种相互作用导致更倾斜的取向，使其中一个苯环更接近Ala47910.51/Ala48010.52。Ala47910.51和Ala48010.52突变为缬氨酸支持了这些预测，BZT的亲和力比JHW 007的亲和力降低更大。总之，我们的数据表明，BZTs显示一个经典的竞争性结合模式与可卡因和多巴胺的结合位点重叠。
Analogues of benztropines (BZTs) are potent inhibitors of the dopamine transporter (DAT) but are less effective than cocaine as behavioral stimulants. As a result, there have been efforts to evaluate these compounds as leads for potential medication for cocaine addiction. Here we use computational modeling together with site-directed mutagenesis to characterize the binding site for BZTs in DAT. Docking into molecular models based on the structure of the bacterial homologue LeuT supported a BZT binding site that overlaps with the substrate binding pocket. In agreement, mutations of residues within the pocket, including Val1523.46* to Ala or Ile, Ser4228.60 to Ala and Asn1573.51 to Cys or Ala, resulted in decreased affinity for BZT and the analog JHW007, as assessed in [3H]dopamine uptake inhibition assays and/or [3H]CFT competition binding assay. A putative polar interaction of one of the phenyl ring fluorine substituents in JHW007 with Asn1573.51 was used as a criterion for determining likely binding poses and establish a structural context for the mutagenesis findings. The analysis positioned the other fluorine substituted phenyl ring of JHW007 in close proximity to Ala47910.51/Ala48010.52 in transmembrane segment (TM) 10. The lack of such an interaction for BZT led to a more tilted orientation, as compared to JHW007, bringing one of the phenyl rings even closer to Ala47910.51/Ala48010.52. Mutation of Ala47910.51 and Ala48010.52 to valines supported these predictions with a larger decrease in the affinity for BZT than for JHW007. Summarized, our data suggest that BZTs display a classical competitive binding mode with binding sites overlapping those of cocaine and dopamine.
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影响因子：
7.3
作者：
Iversen, L
通讯作者：
Iversen, L
影响因子：
7.3
作者：
Agoston, GE;Wu, JH;Newman, AH
通讯作者：
Newman, AH
DOI：
10.1073/pnas.032386299
发表时间：
2002-02-05
影响因子：
11.1
作者：
Loland, CJ;Norregaard, L;Gether, U
通讯作者：
Gether, U
影响因子：
3.6
作者：
Guptaroy, Bipasha;Zhang, Minjia;Gnegy, Margaret E.
通讯作者：
Gnegy, Margaret E.
影响因子：
4.8
作者：
Andersen, Jacob;Taboureau, Olivier;Kristensen, Anders S.
通讯作者：
Kristensen, Anders S.