Mutations affecting the oligomerization interface of G-protein-coupled receptors revealed by a novel de novo protein design framework.
Mutations affecting the oligomerization interface of G-protein-coupled receptors revealed by a novel de novo protein design framework.
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DOI：
10.1529/biophysj.107.117622
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发表时间：
2008-04
影响因子：
3.4
通讯作者：
Martin S. Taylor;H. K. Fung;R. Rajgaria;M. Filizola;H. Weinstein;C. Floudas
中科院分区：
文献类型：
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作者：
Martin S. Taylor;H. K. Fung;R. Rajgaria;M. Filizola;H. Weinstein;C. Floudas
特定的功能和药理学特性最近被归因于G蛋白偶联受体（GPCR）二聚体/寡聚体。由于两个相同或两个不同的GPCR单体的缔合似乎是引发受体功能所必需的，因此有必要了解这种相互作用的确切性质。我们在这里提出了一种新的方法从头蛋白质的设计和它的应用预测的突变，可以稳定或不稳定的GPCR二聚体，同时保持单体的天然折叠。为了测试这种新方法的有效性，血型糖蛋白A的单跨膜结构域的二聚体被用作模型系统。从诱变的螺旋-螺旋界面的实验数据进行比较，在该接口的计算预测，模型的结果被发现是与实验结果一致。一个灵活的模板开发的视紫红质同源二聚体在原子分辨率和用于预测集的三个和五个突变。结果发现，在8个案例研究中是一致的，在每个位置的有利突变。发现预测为在二聚化界面处最具破坏性的突变集合对柔性模板的特异性低于预测为较不具破坏性的集合。
Specific functional and pharmacological properties have recently been ascribed to G-protein-coupled receptor (GPCR) dimers/oligomers. Because the association of two identical or two distinct GPCR monomers seems to be required to elicit receptor function, it is necessary to understand the exact nature of this interaction. We present here a novel method for de novo protein design and its application to the prediction of mutations that can stabilize or destabilize a GPCR dimer while maintaining the monomer's native fold. To test the efficacy of this new method, the dimer of the single-spanned transmembrane domain of glycophorin A was used as a model system. Experimental data from mutagenesis of the helix-helix interface are compared with computational predictions at that interface, and the model's results are found to be consistent with the experimental findings. A flexible template was developed for the rhodopsin homodimer at atomic resolution and used to predict sets of three and five mutations. The results are found to be consistent across eight case studies, with favored mutations at each position. Mutation sets predicted to be the most disruptive at the dimerization interface are found to be less specific to the flexible template than sets predicted to be less disruptive.