Piraner Dan I, Wu Yan, Shapiro Mikhail G
ACS Synth Biol. 2019 Oct 18;8(10):2256-2262. doi: 10.1021/acssynbio.9b00275. Epub 2019 Sep 12.
Protein-protein interactions and protein localization are essential mechanisms of cellular signal transduction. The ability to externally control such interactions using chemical and optogenetic methods has facilitated biological research and provided components for the engineering of cell-based therapies and materials. However, chemical and optical methods are limited in their ability to provide spatiotemporal specificity in light-scattering tissues. To overcome these limitations, we present "thermomers", modular protein dimerization domains controlled with temperature-a form of energy that can be delivered to cells both globally and locally in a wide variety of and contexts. Thermomers are based on a sharply thermolabile coiled-coil protein, which we engineered to heterodimerize at a tunable transition temperature within the biocompatible range of 37-42 °C. When fused to other proteins, thermomers can reversibly control their association, as demonstrated membrane localization in mammalian cells. This technology enables remote control of intracellular protein-protein interactions with a form of energy that can be delivered with spatiotemporal precision in a wide range of biological, therapeutic, and living material scenarios.
蛋白质-蛋白质相互作用和蛋白质定位是细胞信号转导的基本机制。利用化学和光遗传学方法从外部控制此类相互作用的能力促进了生物学研究，并为基于细胞的疗法和材料工程提供了组件。然而，化学和光学方法在光散射组织中提供时空特异性的能力有限。为了克服这些限制，我们提出了“热体”，即由温度控制的模块化蛋白质二聚化结构域——一种可以在各种不同情况下全局和局部传递到细胞的能量形式。热体基于一种对温度敏感的卷曲螺旋蛋白，我们对其进行了工程改造，使其在37-42°C生物相容性范围内的可调转变温度下异源二聚化。当与其他蛋白质融合时，热体可以可逆地控制它们的结合，如在哺乳动物细胞中的膜定位所示。这项技术能够以一种可以在广泛的生物学、治疗和生物材料场景中以时空精度传递的能量形式远程控制细胞内蛋白质-蛋白质相互作用。
