Department of Molecular and Cellular Biophysics, Max Planck Institute of Biochemistry, 82152, Martinsried, Germany.
School of Chemistry, University of Bristol, Cantock's Close, Bristol, BS8 1TS, UK.
Nat Commun. 2021 Mar 5;12(1):1472. doi: 10.1038/s41467-021-21622-5.
Modules that switch protein-protein interactions on and off are essential to develop synthetic biology; for example, to construct orthogonal signaling pathways, to control artificial protein structures dynamically, and for protein localization in cells or protocells. In nature, the E. coli MinCDE system couples nucleotide-dependent switching of MinD dimerization to membrane targeting to trigger spatiotemporal pattern formation. Here we present a de novo peptide-based molecular switch that toggles reversibly between monomer and dimer in response to phosphorylation and dephosphorylation. In combination with other modules, we construct fusion proteins that couple switching to lipid-membrane targeting by: (i) tethering a 'cargo' molecule reversibly to a permanent membrane 'anchor'; and (ii) creating a 'membrane-avidity switch' that mimics the MinD system but operates by reversible phosphorylation. These minimal, de novo molecular switches have potential applications for introducing dynamic processes into designed and engineered proteins to augment functions in living cells and add functionality to protocells.
模块开关蛋白-蛋白相互作用的和关闭是必不可少的发展合成生物学;例如,构建正交信号通路,动态控制人工蛋白质结构,和蛋白质在细胞或原细胞的定位。在自然界中,大肠杆菌 MinCDE 系统耦合核苷酸依赖的切换 MinD 二聚体的膜靶向引发时空模式形成。在这里,我们提出了一个从头多肽为基础的分子开关,可以在磷酸化和去磷酸化的反应之间可逆地切换单体和二聚体。结合其他模块,我们构建融合蛋白的开关与脂质膜的目标: (i) 可逆地将“货物”分子固定在永久的膜“锚”上;和 (ii) 创建一个“膜亲和力开关”，模拟 MinD 系统，但通过可逆磷酸化作用。这些最小的、从头开始的分子开关在引入动态过程到设计和工程蛋白中具有潜在的应用,以增强活细胞中的功能,并为原细胞添加功能。
