ARC Centre of Excellence in Synthetic Biology, Brisbane, Queensland, Australia.
Centre for Agriculture and the Bioeconomy, Queensland University of Technology, Brisbane, Queensland, Australia.
Nat Nanotechnol. 2023 Nov;18(11):1327-1334. doi: 10.1038/s41565-023-01450-y. Epub 2023 Jul 27.
The construction and assembly of artificial allosteric protein switches into information and energy processing networks connected to both biological and non-biological systems is a central goal of synthetic biology and bionanotechnology. However, designing protein switches with the desired input, output and performance parameters is challenging. Here we use a range of reporter proteins to demonstrate that their chimeras with duplicated receptor domains produce YES gate protein switches with large (up to 9,000-fold) dynamic ranges and fast (minutes) response rates. In such switches, the epistatic interactions between largely independent synthetic allosteric sites result in an OFF state with minimal background noise. We used YES gate protein switches based on β-lactamase to develop quantitative biosensors of therapeutic drugs and protein biomarkers. Furthermore, we demonstrated the reconfiguration of YES gate switches into AND gate switches controlled by two different inputs, and their assembly into signalling networks regulated at multiple nodes.
将人工别构蛋白开关构建和组装到与生物和非生物系统相连的信息和能量处理网络中，是合成生物学和生物纳米技术的核心目标。然而，设计具有所需输入、输出和性能参数的蛋白开关具有挑战性。在这里，我们使用一系列报告蛋白证明，它们与重复受体结构域的嵌合体产生具有大（高达 9000 倍）动态范围和快速（分钟级）响应速率的 YES 门蛋白开关。在这些开关中，在很大程度上独立的人工别构位点之间的上位相互作用导致具有最小背景噪声的 OFF 状态。我们使用基于β-内酰胺酶的 YES 门蛋白开关来开发治疗药物和蛋白质生物标志物的定量生物传感器。此外，我们还证明了 YES 门开关可以重新配置为由两个不同输入控制的 AND 门开关，并将它们组装成在多个节点受到调节的信号网络。
