Computational design of receptor and sensor proteins with novel functions
Computational design of receptor and sensor proteins with novel functions
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DOI：
10.1038/nature01556
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发表时间：
2003-05-08
期刊：
影响因子：
64.8
通讯作者：
Hellinga, HW
中科院分区：
文献类型：
--
作者：
Looger, LL;Dwyer, MA;Hellinga, HW
在分子水平上，蛋白质和配体之间形成复合物是生物过程的基础。因此，操纵配体和蛋白质之间的分子识别对于基础生物学研究非常重要(1)，并且具有许多生物技术应用，包括酶的构建（2-4）、生物传感器（5,6）、遗传电路(7)、信号转导途径(8)和手性分离(9)。结合位点的系统操作仍然是一个主要的挑战。计算设计为工程蛋白质结构和功能提供了巨大的通用性（10）。在这里，我们提出了一种基于结构的计算方法，可以彻底重新设计蛋白质配体结合特异性。该方法构建了高选择性和高亲和力结合三硝基甲苯、l -乳酸或5 -羟色胺的可溶性受体。这些工程受体可以作为新配体的生物传感器；我们还将它们整合到合成细菌信号转导通路中，调节细胞外三硝基甲苯或l -乳酸的基因表达。各种配体和蛋白质的使用表明，对生物分子识别的高度控制已经在计算上建立起来。设计的受体的生物学和生物传感活性说明了计算设计的潜在应用。
The formation of complexes between proteins and ligands is fundamental to biological processes at the molecular level. Manipulation of molecular recognition between ligands and proteins is therefore important for basic biological studies(1) and has many biotechnological applications, including the construction of enzymes(2-4), biosensors(5,6), genetic circuits(7), signal transduction pathways(8) and chiral separations(9). The systematic manipulation of binding sites remains a major challenge. Computational design offers enormous generality for engineering protein structure and function(10). Here we present a structure-based computational method that can drastically redesign protein ligand-binding specificities. This method was used to construct soluble receptors that bind trinitrotoluene, L-lactate or serotonin with high selectivity and affinity. These engineered receptors can function as biosensors for their new ligands; we also incorporated them into synthetic bacterial signal transduction pathways, regulating gene expression in response to extracellular trinitrotoluene or L-lactate. The use of various ligands and proteins shows that a high degree of control over biomolecular recognition has been established computationally. The biological and biosensing activities of the designed receptors illustrate potential applications of computational design.