理性设计别构调控的蛋白质活性开关

蛋白质的别构效应使得蛋白质结构中相互远离的位点产生功能上的耦合。别构效应可以调控细胞内蛋白质活性的高低，它对信号传导、新陈代谢、基因调控网络等细胞功能的调节具有重要的意义。将蛋白质的别构位点与理性设计的分子开关相融合，我们可以使蛋白质响应外界信号（光或小分子），通过别构位点的构象变化 “打开”与“关闭”蛋白质的功能。本项目以Rho蛋白质家族为研究对象，结合别构位点的计算预测方法与实验测量的蛋白质动力学数据，理性地设计光控的GEF、GTPase、Kinase等蛋白质，实现对细胞运动的人工控制。本项目使用人工设计的蛋白质，深入地阐明蛋白质结构内残基-残基关联网络与别构效应的关系，为理性地开发蛋白活性开关和人工调节的功能模块提供可靠的手段。

The allosteric effects enable functional coupling between distant sites within protein structures. It can regulate the intrinsic activity of cellular proteins; hence, allosteric effects are import for cellular functions including signaling transduction、metabolism、genetic networks. Rationally designed molecular switches make possible of engineered proteins responding to external signals (light-sensitive or chemical-sensitive) and altering their functions. Combining the allosteric sites and molecular switches, we can “turn on” or “turn off” protein activities in real time and in situ. In the current project, we will rationally design light-controlled GEF、GTPase、and Kinase, all belonging to the Rho family proteins and important for cell movement. We will combine computational methods to predict allosteric sites and experimental methods to measure protein dynamics, to elucidate the relationship between allosteric effects and residue-residue correlation networks within designed proteins. Eventually, we can achieve modulated morphodynamics of living cells and provide robust methods for engineering allosterically-regulated protein switches and functional modules.