Super-complexes of adhesion GPCRs and neural guidance receptors.
Super-complexes of adhesion GPCRs and neural guidance receptors.
复制标题
DOI：
10.1038/ncomms11184
复制
发表时间：
2016-04-19
影响因子：
16.6
通讯作者：
Seiradake E
中科院分区：
文献类型：
--
作者：
Jackson VA;Mehmood S;Chavent M;Roversi P;Carrasquero M;Del Toro D;Seyit-Bremer G;Ranaivoson FM;Comoletti D;Sansom MS;Robinson CV;Klein R;Seiradake E
亲Latrophilin粘附-GPCR（Lphn 1 -3或ADGRL 1 -3）和Unc 5细胞导向受体（Unc 5A-D）与FLRT蛋白（FLRT 1 -3）相互作用，从而分别促进细胞粘附和排斥。这三种蛋白质如何相互作用并同时发挥作用还知之甚少。我们表明，Unc 5D相互作用FLRT 2顺式，控制细胞粘附响应外部提出的Lphn 3。三种蛋白质的胞外域协同结合。由细胞外结构域形成的三元复合物的晶体结构显示，Lphn 3在与FLRT 2：Unc 5结合时二聚化，导致1：1：2的化学计量（FLRT 2：Unc 5D：Lphn 3）。该1：1：2复合物进一步二聚化以形成更大的“超复合物”（2：2：4），使用Unc 5D TSP 1结构域中先前未描述的结合基序。分子动力学模拟、定点突变和质谱分析证明了这些复合物的稳定性和分子性质。我们的数据表明受体通过形成不同的上下文依赖性高阶复合物来增加其功能库。 已知FLRT蛋白与Lphns和Unc 5相互作用，分别介导细胞粘附和排斥。在这里，作者使用晶体学，天然质谱，分子动力学模拟和基于细胞的测定来证明这三种蛋白质形成了大型的超级复合物，其功能与较小的子复合物不同。
Latrophilin adhesion-GPCRs (Lphn1–3 or ADGRL1–3) and Unc5 cell guidance receptors (Unc5A–D) interact with FLRT proteins (FLRT1–3), thereby promoting cell adhesion and repulsion, respectively. How the three proteins interact and function simultaneously is poorly understood. We show that Unc5D interacts with FLRT2 in cis, controlling cell adhesion in response to externally presented Lphn3. The ectodomains of the three proteins bind cooperatively. Crystal structures of the ternary complex formed by the extracellular domains reveal that Lphn3 dimerizes when bound to FLRT2:Unc5, resulting in a stoichiometry of 1:1:2 (FLRT2:Unc5D:Lphn3). This 1:1:2 complex further dimerizes to form a larger ‘super-complex' (2:2:4), using a previously undescribed binding motif in the Unc5D TSP1 domain. Molecular dynamics simulations, point-directed mutagenesis and mass spectrometry demonstrate the stability and molecular properties of these complexes. Our data exemplify how receptors increase their functional repertoire by forming different context-dependent higher-order complexes. FLRT proteins are known to interact with Lphns and Unc5s, mediating cell adhesion and repulsion respectively. Here the authors use crystallography, native mass spectrometry, molecular dynamics simulations and cell-based assays to show that these three proteins form large super-complexes with functions distinct from their smaller subcomplexes.