The conformation of neurotensin bound to its G protein-coupled receptor
The conformation of neurotensin bound to its G protein-coupled receptor
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DOI：
10.1073/pnas.1834523100
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发表时间：
2003-09-16
影响因子：
11.1
通讯作者：
Baldus, M
中科院分区：
文献类型：
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作者：
Luca, S;White, JF;Baldus, M
G蛋白偶联受体（GPCR）介导嗅觉、光、味觉和疼痛的感知。它们参与信号识别和细胞通讯，是药物开发的一些最重要的靶点。由于目前没有直接的结构信息结合到GPCR的高亲和力配体，合理的药物设计是有限的计算预测结合诱变实验。在这里，我们提出了一个高亲和力的肽激动剂（神经降压素，NT）绑定到其GPCR NTS-1的构象，通过直接的结构方法确定。在大肠杆菌中表达功能性受体，通过使用优化的程序以毫克量纯化，随后重构成脂质囊泡。定制固态NMR实验以允许明确检测与功能性NTS-1复合的微克量的C-13、N-15标记的NT（8-13）。NMR数据与在不存在受体的情况下配体的无序状态一致。在受体结合时，肽经历线性重排，采用β-链构象。我们的研究结果提供了一个可行的结构模板，为进一步的药理学研究。
G protein-coupled receptors (GPCRs) mediate the perception of smell, light, taste, and pain. They are involved in signal recognition and cell communication and are some of the most important targets for drug development. Because currently no direct structural information on high-affinity ligands bound to GPCRs is available, rational drug design is limited to computational prediction combined with mutagenesis experiments. Here, we present the conformation of a high-affinity peptide agonist (neurotensin, NT) bound to its GPCR NTS-1, determined by direct structural methods. Functional receptors were expressed in Escherichia coli, purified in milligram amounts by using optimized procedures, and subsequently reconstituted into lipid vesicles. Solid-state NMR experiments were tailored to allow for the unequivocal detection of microgram quantities of C-13,N-15-labeled NT(8-13) in complex with functional NTS-1. The NMR data are consistent with a disordered state of the ligand in the absence of receptor. Upon receptor binding, the peptide undergoes a linear rearrangement, adopting a beta-strand conformation. Our results provide a viable structural template for further pharmacological investigations.