Properties of the N‐terminal domains from Y receptors probed by NMR spectroscopy
Properties of the N‐terminal domains from Y receptors probed by NMR spectroscopy
复制标题
通过 NMR 波谱探测 Y 受体 N 端结构域的特性
DOI：
10.1002/psc.1102
复制
发表时间：
2009
影响因子：
2.1
通讯作者：
O. Zerbe
中科院分区：
文献类型：
--
作者：
C. Zou;Sowmini Kumaran;R. Walser;O. Zerbe
来自NPY家族的神经激素与其受体（所谓的Y受体，属于G蛋白偶联受体超家族1b）的结合可能包括与受体N末端结构域的瞬时结合。因此，我们研究了来自Y1、Y2、Y 4和Y 5受体亚型（N-Y1、N-Y2、N-Y 4、N-Y 5）的N末端结构域的结构特征。我们为它们的重组表达开发了有效的策略。N-Y 4和N-Y1表达为不溶性融合物以促进积累到包涵体中，而N-Y2和N-Y 5表达为可溶性融合蛋白。所有N-末端结构域在水性缓冲液中是完全柔性的。在磷脂胶束的存在下，多肽内的一些伸展采用螺旋构象，但这些太不稳定而不能详细表征。使用化学位移作图技术，NPY、肽YY（PYY）和胰多肽（PP）（神经激素家族的三个成员，是Y受体的天然配体）与N-Y1、N-Y2和N-Y 5的相互作用在所有情况下都显示出化学位移变化，其中在存在和不存在磷脂胶束的情况下，PP与N-Y1或N-Y 5相互作用的值最大。然而，相互作用的强度通常较弱，数据也指向非特异性接触。以前，在N-Y 4与PP相互作用的情况下，接触被证明是静电性质的。这项工作表明，肽与N末端结构域的缔合通常可能是其结合轨迹的一部分。版权所有© 2008欧洲肽协会和约翰威利父子有限公司。
Binding of neurohormones from the NPY family to their receptors, the so‐called Y receptors, that belong to the superfamily 1b of G‐protein coupled receptors might include transient binding to the N‐terminal domains of the receptors. Accordingly, we have studied structural features of the N‐terminal domains from the Y1, Y2, Y4, and Y5 receptor subtypes (N‐Y1, N‐Y2, N‐Y4, N‐Y5). We developed efficient strategies for their recombinant expression. N‐Y4 and N‐Y1 were expressed as insoluble fusions to enforce accumulation into inclusion bodies, whereas N‐Y2 and N‐Y5 were expressed as soluble fusion proteins. All N‐terminal domains are fully flexible in aqueous buffer. In the presence of phospholipid micelles some stretches within the polypeptides adopt helical conformations, but these are too unstable to be characterized in detail. Using chemical shift mapping techniques, interactions of NPY, peptide YY (PYY), and pancreatic polypeptide (PP), the three members of the neurohormone family that are the Y receptors' natural ligands, with N‐Y1, N‐Y2, and N‐Y5 revealed chemical shift changes in all cases, with the largest values being encountered for PP interacting with N‐Y1 or N‐Y5 both in the presence and in the absence of phospholipid micelles. The strength of the interactions, however, is generally weak, and the data also point to nonspecific contacts. Previously, in case of the interaction of N‐Y4 with PP, the contacts were shown to be electrostatic in nature. This work indicates that association of the peptides with the N‐terminal domains may generally be part of their binding trajectory. Copyright © 2008 European Peptide Society and John Wiley & Sons, Ltd.
影响因子：
1.6
作者：
Mohanty, AK;Simmons, CR;Wiener, MC
通讯作者：
Wiener, MC
影响因子：
56.9
作者：
Palczewski, K;Kumasaka, T;Miyano, M
通讯作者：
Miyano, M