Department of Molecular Biology and Nanobiotechnology , National Institute of Chemistry , Hajdrihova ulica 19 , 1001 Ljubljana , Slovenia.
Biomedicine Doctoral Program , University of Ljubljana , Vrazov trg 2 , 1000 Ljubljana , Slovenia.
ACS Synth Biol. 2020 Feb 21;9(2):316-328. doi: 10.1021/acssynbio.9b00340. Epub 2020 Feb 11.
Lipid membranes are becoming increasingly popular in synthetic biology due to their biophysical properties and crucial role in communication between different compartments. Several alluring protein-membrane sensors have already been developed, whereas protein logic gates designs on membrane-embedded proteins are very limited. Here we demonstrate the construction of a two-level protein-membrane logic gate with an OR-AND logic. The system consists of an engineered pH-dependent pore-forming protein listeriolysin O and its DARPin-based inhibitor, conjugated to a lipid vesicle membrane. The gate responds to low pH and removal of the inhibitor from the membrane either by switching to a reducing environment, protease cleavage, or any other signal depending on the conjugation chemistry used for inhibitor attachment to the membrane. This unique protein logic gate vesicle system advances generic sensing and actuator platforms used in synthetic biology and could be utilized in drug delivery.
由于脂质膜具有生物物理特性及其在不同隔室之间通讯中的关键作用，因此在合成生物学中越来越受欢迎。已经开发出了几种诱人的蛋白-膜传感器，而在膜嵌入蛋白上设计蛋白逻辑门则非常有限。在这里，我们展示了一个具有 OR-AND 逻辑的两级蛋白-膜逻辑门的构建。该系统由工程化的 pH 依赖性孔形成蛋白溶菌酶 O 及其基于 DARPin 的抑制剂组成，连接到脂质囊泡膜上。该门在低 pH 值下以及从膜上除去抑制剂时做出响应，通过切换到还原环境、蛋白酶切割或任何其他信号，具体取决于用于将抑制剂附着到膜上的缀合化学。这种独特的蛋白逻辑门囊泡系统推进了在合成生物学中使用的通用传感和执行器平台，并且可以用于药物输送。
