Probing the environment of neurotensin whilst bound to the neurotensin receptor by solid state NMR
Probing the environment of neurotensin whilst bound to the neurotensin receptor by solid state NMR
复制标题
通过固态 NMR 探测神经降压素与神经降压素受体结合的环境
DOI：
10.1016/s0014-5793(02)02656-x
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发表时间：
2002
期刊：
影响因子：
3.5
通讯作者：
A. Watts
中科院分区：
文献类型：
--
作者：
P. Williamson;S. Bains;C. Chung;R. Cooke;A. Watts
已经通过核磁共振（NMR）观察到神经降压素的功能活性类似物神经降压素（8-13）与大鼠神经降压素受体的激动剂结合位点结合。通过应用缓慢魔角样品旋转和高功率质子去耦，在碳-13光谱中获得足够的分辨率和灵敏度，以允许分配由均匀碳-13/氮-15-标记的神经降压素（8-13）产生的许多侧链共振，同时与神经降压素受体结合。在神经降压素（8-13）与受体结合后观察到碳-13化学位移的显著扰动。最重要的是，在神经降压素的羧基末端和酪氨酸侧链中观察到显著的位移（8-13），表明这些位点在神经降压素与神经降压素受体上的激动剂结合位点的相互作用中是重要的。相反，没有观察到干扰的精氨酸侧链的胍基内的碳-13位点，表明与受体结合位点的相互作用很小，或周围的氮原子屏蔽局部环境。这些NMR观测进一步支持以前的结构-活性研究，定点诱变和建模研究的激动剂结合位点的神经降压素受体，从相同的特定残基的NMR扰动被观察到的神经降压素受体激活神经降压素是重要的。
A functionally active analogue of neurotensin, neurotensin(8–13), has been observed whilst bound to the agonist-binding site of the rat neurotensin receptor by nuclear magnetic resonance (NMR). Through the application of slow magic angle sample spinning and high-power proton decoupling, sufficient resolution and sensitivity were obtained in the carbon-13 spectrum to allow an assignment of many of the side chain resonances arising from uniformly carbon-13/nitrogen-15-labelled neurotensin(8–13) whilst bound to the neurotensin receptor. Significant perturbations in carbon-13 chemical shift were observed upon the binding of the neurotensin(8–13) to the receptor. Most importantly significant shifts were observed in both the carboxy terminus and tyrosine side chain of the neurotensin(8–13), suggesting that these sites are important in the interaction of the neurotensin with the agonist-binding site on the neurotensin receptor. Conversely, no perturbations were observed for the carbon-13 sites within the guanidinium groups of the arginine side chains, indicating little interaction with the receptor-binding site, or a shielding of the local environment by the surrounding nitrogen atoms. These NMR observations lend further support to previous structure–activity studies, site-directed mutagenesis and modelling studies of the agonist-binding site of the neurotensin receptor, from which the same specific residues for which NMR perturbations were observed are important for neurotensin receptor activation by neurotensin.