Engineered allosteric regulation of protein activity provides significant advantages for the development of robust and broadly applicable tools. However, the application of allosteric switches in optogenetics has been scarce and suffers from critical limitations. Here, we report an optogenetic approach that utilizes an engineered Light-Regulated (LightR) allosteric switch module to achieve tight spatiotemporal control of enzymatic activity. Using the tyrosine kinase Src as a model, we demonstrate efficient regulation of the kinase and identify temporally distinct signaling responses ranging from seconds to minutes. LightR-Src off-kinetics can be tuned by modulating the LightR photoconversion cycle. A fast cycling variant enables the stimulation of transient pulses and local regulation of activity in a selected region of a cell. The design of the LightR module ensures broad applicability of the tool, as we demonstrate by achieving light-mediated regulation of Abl and bRaf kinases as well as Cre recombinase.

中文翻译：

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光调节变构开关能够对酶活性进行时间和亚细胞控制


蛋白质活性的工程变构调节为开发稳健且广泛适用的工具提供了显着的优势。然而，变构开关在光遗传学中的应用很少，并且受到严重限制。在这里，我们报告了一种光遗传学方法，该方法利用工程化的光调节（LightR）变构开关模块来实现酶活性的严格时空控制。使用酪氨酸激酶 Src 作为模型，我们证明了激酶的有效调节，并识别了从几秒到几分钟的时间上不同的信号反应。 LightR-Src 的离动力学可以通过调节 LightR 光转换周期来调节。快速循环变体能够刺激瞬态脉冲并局部调节细胞选定区域的活动。 LightR 模块的设计确保了该工具的广泛适用性，正如我们通过实现 Abl 和 bRaf 激酶以及 Cre 重组酶的光介导调节所证明的那样。