Surface Plasmon Resonance Biosensor Based Fragment Screening Using Acetylcholine Binding Protein Identifies Ligand Efficiency Hot Spots (LE Hot Spots) by Deconstruction of Nicotinic Acetylcholine Receptor α7 Ligands
Surface Plasmon Resonance Biosensor Based Fragment Screening Using Acetylcholine Binding Protein Identifies Ligand Efficiency Hot Spots (LE Hot Spots) by Deconstruction of Nicotinic Acetylcholine Receptor α7 Ligands
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DOI：
10.1021/jm100834y
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发表时间：
2010-10-14
影响因子：
7.3
通讯作者：
de Esch, Iwan J. P.
中科院分区：
文献类型：
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作者：
de Kloe, Gerdien E.;Retra, Kim;de Esch, Iwan J. P.
可溶乙酰胆碱结合蛋白(AChBP)是烟碱型乙酰胆碱受体(NAChR)配体结合域的同源物。为了指导未来利用表面等离子体共振(SPR)生物传感器技术进行片段筛选，作为一种无标记、直接结合的生物物理筛选方法，基于一组含有具有纳米分子亲和力的α7 nAChR选择性奎宁核苷配体的解构，构建了一个聚焦片段文库。用SPR生物传感器实验评价了这些片段和母体化合物与AChBP的相互作用特性。从这种直接结合分析获得的数据与参考放射性配基置换分析的数据具有很好的相关性。文库中不同(结构)片段组的配基效率与与结合口袋的不同区域的结合相关，从而识别配基效率热点(LE热点)。这些热点可用于在大规模片段文库筛选中识别最有希望的命中片段。
The soluble acetylcholine binding protein (AChBP) is a homologue of the ligand-binding domain of the nicotinic acetylcholine receptors (nAChR). To guide future fragment-screening using surface plasmon resonance (SPR) biosensor technology as a label-free, direct binding, biophysical screening assay, a focused fragment library was generated based on deconstruction of a set of alpha 7 nAChR selective quinuclidine containing ligands with nanomolar affinities. The interaction characteristics of the fragments and the parent compounds with AChBP) were evaluated using an SPR biosensor assay. The data obtained from this direct binding assay correlated well with data from the reference radioligand displacement assay. Ligand efficiencies for different (structural) groups of fragments in the library were correlated to binding with distinct regions of the binding pocket, thereby identifying ligand efficiency hot spots (LE hot spots). These hot spots can be used to identity the most promising hit fragments in a large scale fragment library screen.