Ribeiro Lucas F, Warren Tiana D, Ostermeier Marc
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles St, Baltimore, MD, 21218, USA.
Methods Mol Biol. 2017;1596:43-55. doi: 10.1007/978-1-4939-6940-1_3.
A protein switch is a protein that changes between inactive ("off") and active ("on") states in response to a biomolecule or physical signal. These switches can be created by fusing two domains in such a way that the activity of the output domain is regulated by the input domain's recognition of an input signal (such as the binding of a molecule, recognition of light). Here, we describe several methods for randomly fusing two domains to create domain insertion libraries from which protein switches can be identified by selections and/or screens.
蛋白质开关是一种蛋白质，它会根据生物分子或物理信号在非活性（“关闭”）和活性（“开启”）状态之间切换。这些开关可以通过将两个结构域融合来创建，使得输出结构域的活性由输入结构域对输入信号的识别（例如分子的结合、光的识别）来调节。在这里，我们描述了几种随机融合两个结构域以创建结构域插入文库的方法，通过筛选和/或选择可以从该文库中鉴定出蛋白质开关。
