Low-stringency selection of TEM1 for BLIP shows interface plasticity and selection for faster binders
Low-stringency selection of TEM1 for BLIP shows interface plasticity and selection for faster binders
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DOI：
10.1073/pnas.1613122113
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发表时间：
2016-12-27
影响因子：
11.1
通讯作者：
Schreiber, Gideon
中科院分区：
文献类型：
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作者：
Cohen-Khait, Ruth;Schreiber, Gideon
蛋白质-蛋白质相互作用通过蛋白质表面上明确定义的界面发生。而同源接口的位置是保守的，它们的组成不同，这表明多种解决方案可能支持高亲和力结合。在这项研究中，我们研究了可塑性的界面TEM 1 β-内酰胺酶与其蛋白质抑制剂BLIP通过低严格性选择的随机TEM 1库使用酵母表面展示。我们的研究结果表明，大多数界面残基可以突变，而不会损失结合亲和力，蛋白质稳定性，或酶活性，这表明在界面组合物的可塑性支持高亲和力结合。有趣的是，许多选定的突变促进了更快的关联。通过将库配体孵育时间大幅减少至30秒来实现对更快结合剂的进一步选择。本文提出的预平衡选择是一种特异性选择快速缔合蛋白质复合物的新方法。
Protein-protein interactions occur via well-defined interfaces on the protein surface. Whereas the location of homologous interfaces is conserved, their composition varies, suggesting that multiple solutions may support high-affinity binding. In this study, we examined the plasticity of the interface of TEM1 beta-lactamase with its protein inhibitor BLIP by low-stringency selection of a random TEM1 library using yeast surface display. Our results show that most interfacial residues could be mutated without a loss in binding affinity, protein stability, or enzymatic activity, suggesting plasticity in the interface composition supporting high-affinity binding. Interestingly, many of the selected mutations promoted faster association. Further selection for faster binders was achieved by drastically decreasing the libraryligand incubation time to 30 s. Preequilibrium selection as suggested here is a novel methodology for specifically selecting faster-associating protein complexes.