Binding between a distal C-terminus fragment of cannabinoid receptor 1 and arrestin-2.
Binding between a distal C-terminus fragment of cannabinoid receptor 1 and arrestin-2.
复制标题
DOI：
10.1021/bi1018144
复制
发表时间：
2011-03-29
期刊：
影响因子：
2.9
通讯作者：
Pavlopoulos S
中科院分区：
文献类型：
--
作者：
Singh SN;Bakshi K;Mercier RW;Makriyannis A;Pavlopoulos S
g蛋白偶联受体的内化是通过c端磷酸化介导的，随后与胞质蛋白阻滞蛋白结合。为了探索可能在大麻素受体1 （CB1）内化中发挥作用的结构因素，我们利用了CB1远端c端衍生的磷酸化肽（CB15P454-473）。用等温滴定量热法和核磁共振波谱法比较了肽与人类arrestin-2 （wt-arr21-418）形成的复合物与截断的arrestin-2突变体（tr-arr21-382）形成的复合物。五磷酸肽CB15P454-473无论是与全长阻滞蛋白2还是其截短突变体结合，都采用螺旋环构象。这种结构类似于七肽-磷酸肽的结构，模仿视紫红质c -尾的远端片段（Rh7P330-348），与视觉阻滞蛋白结合，表明这种结构具有功能意义。等温滴定量热法（ITC）实验表明，CB15P454-473肽与tr-arr21-382的结合亲和力高于与全长wt-arr21-418的结合亲和力。由于观察到的结合肽的结构在两种情况下都是相似的，我们将增加的亲和力归因于截断的抑制蛋白结构的n结构域上更暴露的结合位点。利用数据驱动的HADDOCK对接程序，从结合的磷酸肽转移的nOe数据用于预测描述与抑制蛋白相互作用的模型。arrestin-2的截断为该蛋白极核中带正电的残基与CB15P454-473肽环中存在的磷酸盐相互作用提供了空间。
Internalization of G-protein coupled receptors is mediated by phosphorylation of the C-terminus, followed by binding with the cytosolic protein arrestin. To explore structural factors that may play a role in internalization of cannabinoid receptor 1 (CB1), we utilize a phosphorylated peptide derived from the distal C-terminus of CB1 (CB15P454-473). Complexes formed between the peptide and human arrestin-2 (wt-arr21-418) were compared to those formed with a truncated arrestin-2 mutant (tr-arr21-382) using isothermal titration calorimetry and nuclear magnetic resonance spectroscopy. The penta-phosphopeptide CB15P454-473 adopts a helix-loop conformation, whether binding to full-length arrestin-2 or its truncated mutant. This structure is similar to that of a hepta-phosphopeptide, mimicking the distal segment of the rhodopsin C-tail (Rh7P330-348), binding to visual arrestin, suggesting that this adopted structure bears functional significance. Isothermal titration calorimetry (ITC) experiments show that the CB15P454-473 peptide binds to tr-arr21-382 with higher affinity than to the full-length wt-arr21-418. As the observed structure of the bound peptides is similar in either case, we attribute the increased affinity to a more exposed binding site on the N-domain of the truncated arrestin construct. The transferred nOe data from the bound phosphopeptides are used to predict a model describing the interaction with arrestin, using the data driven HADDOCK docking program. The truncation of arrestin-2 provides scope for positively charged residues in the polar core of the protein to interact with phosphates present in the loop of the CB15P454-473 peptide.
登录
查看更多内容
影响因子：
11.4
作者：
CLORE, GM;NILGES, M;GRONENBORN, AM
通讯作者：
GRONENBORN, AM
影响因子：
4.8
作者：
Anavi-Goffer, Sharon;Fleischer, Daniel;Abood, Mary E.
通讯作者：
Abood, Mary E.
影响因子：
5.6
作者：
CLORE, GM;BRUNGER, AT;GRONENBORN, AM
通讯作者：
GRONENBORN, AM
通过滥交磷酸化依赖性机制调节CB1大麻素受体内在化。
影响因子：
4.7
作者：
Daigle TL;Kwok ML;Mackie K
通讯作者：
Mackie K
影响因子：
4.8
作者：
GUREVICH, VV;BENOVIC, JL
通讯作者：
BENOVIC, JL