Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template.
Generation of candidate ligands for nicotinic acetylcholine receptors via in situ click chemistry with a soluble acetylcholine binding protein template.
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DOI：
10.1021/ja3001858
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发表时间：
2012-04-18
影响因子：
15
通讯作者：
Fokin, Valery V.
中科院分区：
文献类型：
--
作者：
Grimster, Neil P.;Stump, Bernhard;Fotsing, Joseph R.;Weide, Timo;Talley, Todd T.;Yamauchi, John G.;Nemecz, Akos;Kim, Choel;Ho, Kwok-Yiu;Sharpless, K. Barry;Taylor, Palmer;Fokin, Valery V.
烟碱型乙酰胆碱受体（nAChRs）广泛存在于中枢和外周神经系统中，负责介导重要的生理功能。作为Cys环配体门控离子通道家族的成员，神经元nA-ChR是五聚体，由α（α2至α10）和β（β2至β4）亚基的各种排列组成，形成功能性异聚体或同聚体受体。nAChR亚基组成的多样性使特定亚型的选择性配体的开发复杂化，因为五个结合位点位于亚基界面。乙酰胆碱结合蛋白（AChBP）是软体动物分泌的一种可溶性胞外结构域同源物，可作为nAChRs的一般结构替代物。在这项工作中，同源AChBPs从AChBPs和AChBPs蜗牛被用来作为原位模板，用于生成新的和有效的配体，选择性地结合到这些蛋白质。叠氮化物和炔之间的环加成反应生成稳定的1，2，3-三唑产生的铅。AChBP模板上三唑形成的程度与三唑产物在烟碱配体结合位点的亲和力相关。而不是在原位蛋白质模板的叠氮化物-炔环加成反应发生在一个本地化的，隔离的酶活性中心，如前所示，我们证明，原位反应可以发生在亚基的低聚蛋白质的接口，因此可以被用作一种工具，用于识别新的候选nAChR配体。一个在原位形成的三唑-乙酰胆碱BP复合物的晶体结构显示结合姿态和分子决定因素的相互作用预测已知的激动剂和拮抗剂的结构。因此，具有受体的原位模板的点击化学方法为产生配体门控离子通道的候选激动剂和拮抗剂提供了新的合成途径。
Nicotinic acetylcholine receptors (nAChRs), being responsible for mediating key physiological functions, are ubiquitous in the central and peripheral nervous systems. As members of the Cys loop ligand-gated ion channel family, neuronal nA-ChRs are pentameric, composed of various permutations of α (α2 to α10) and β (β2 to β4) subunits forming functional heteromeric or homomeric receptors. Diversity in nAChR subunit composition complicates development of selective ligands for specific subtypes, since the five binding sites reside at the subunit interfaces. The acetylcholine binding protein (AChBP), a soluble extracellular domain homologue secreted by mollusks, serves as a general structural surrogate for the nAChRs. In this work, homomeric AChBPs from Lymnaea and Aplysia snails were used as in situ templates for the generation of novel and potent ligands that selectively bind to these proteins. The cycloaddition reaction between building block azides and alkynes to form stable 1,2,3-triazoles generated the leads. The extent of triazole formation on the AChBP template correlated with the affinity of the triazole product at the nicotinic ligand binding site. Instead of the in situ protein-templated azide-alkyne cycloaddition reaction occurring at a localized, sequestered enzyme active center as previously shown, we demonstrate that the in situ reaction can take place at subunit interfaces of an oligomeric protein and can thus be used as a tool for identification of novel candidate nAChR ligands. The crystal structure of one of the in situ formed triazole–AChBP complexes shows binding poses and molecular determinants of interactions predicted from structures of known agonists and antagonists. Hence, the click chemistry approach with an in situ template of a receptor provides a novel synthetic avenue for generating candidate agonists and antagonists for ligand-gated ion channels.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
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通讯作者：
Cowtan, K
影响因子：
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作者：
Horne, WS;Stout, CD;Ghadiri, MR
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影响因子：
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作者：
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影响因子：
11.4
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Hansen, SB;Sulzenbacher, G;Bourne, Y
通讯作者：
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影响因子：
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作者：
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