Modulating protein activity using tethered ligands with mutually exclusive binding sites
Modulating protein activity using tethered ligands with mutually exclusive binding sites
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DOI：
10.1038/ncomms8830
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发表时间：
2015-07-01
影响因子：
16.6
通讯作者：
Johnsson, Kai
中科院分区：
文献类型：
--
作者：
Schena, Alberto;Griss, Rudolf;Johnsson, Kai
设计其活性可以由不相关的效应分子开启和关闭的蛋白质的可能性将使其在从生物传感到合成生物学的各种研究领域中的应用成为可能。我们在这里描述了一种响应特定效应物浓度调节蛋白质活性的一般方法。该方法基于具有两个互斥结合位点的合成配体，一个用于感兴趣的蛋白质，一个用于效应物。将这样的配体拴系到感兴趣的蛋白质导致分子内配体-蛋白质相互作用，其可以通过效应物的存在而被破坏。具体而言，我们引入了另一种蛋白质，人类碳酸酐酶，其活性可以控制蛋白质或小分子在体外和活细胞，和新的荧光和生物发光生物传感器控制的荧光素酶。
The possibility to design proteins whose activities can be switched on and off by unrelated effector molecules would enable applications in various research areas, ranging from biosensing to synthetic biology. We describe here a general method to modulate the activity of a protein in response to the concentration of a specific effector. The approach is based on synthetic ligands that possess two mutually exclusive binding sites, one for the protein of interest and one for the effector. Tethering such a ligand to the protein of interest results in an intramolecular ligand-protein interaction that can be disrupted through the presence of the effector. Specifically, we introduce a luciferase controlled by another protein, a human carbonic anhydrase whose activity can be controlled by proteins or small molecules in vitro and on living cells, and novel fluorescent and bioluminescent biosensors.