DWI-Leibniz Institute for Interactive Materials, Forckenbeckstr. 50, 52074 Aachen, Germany.
Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Worringerweg 2, 52074 Aachen, Germany.
J Am Chem Soc. 2024 Oct 2;146(39):26947-26956. doi: 10.1021/jacs.4c08207. Epub 2024 Sep 18.
Utilizing ultrasound as an external stimulus to remotely modulate the activity of proteins is an important aspect of sonopharmacology and establishes the basis for the emerging field of sonogenetics. Here, we describe an ultrasound-responsive protein splicing system that enables spatiotemporal control of split-intein-mediated protein ligation. The system utilizes engineered split inteins that are caged and can be activated by thrombin released from a high molar mass DNA-based carrier under focused ultrasound sonication. This approach represents a general method for controlling the functions of proteins of interest by ultrasound, as demonstrated here by the controlled synthesis of the superfolder green fluorescence protein (GFP) and calcitonin. Furthermore, calcitonin receptor-mediated signal transduction in cells was triggered by this system in vitro without harming cell viability. By expanding the sonogenetic toolbox with protein splicing technologies, this study provides a possible pathway to deploy ultrasound for remotely controlling a variety of protein functions in deep tissue in the future.
利用超声作为外部刺激来远程调节蛋白质的活性是声药理学的一个重要方面，为新兴的声遗传学领域奠定了基础。在这里，我们描述了一种超声响应的蛋白质剪接系统，该系统能够实现分裂内含肽介导的蛋白质连接的时空控制。该系统利用了被笼蔽的工程化分裂内含肽，在聚焦超声处理下，从高摩尔质量基于 DNA 的载体中释放出的凝血酶可以激活这些内含肽。正如本文通过对超折叠绿色荧光蛋白（GFP）和降钙素的可控合成所展示的那样，这种方法代表了一种通过超声控制感兴趣的蛋白质功能的通用方法。此外，该系统在体外还触发了降钙素受体介导的细胞信号转导，而不会损害细胞活力。通过将蛋白质剪接技术扩展到声遗传学工具包中，本研究为未来在深部组织中利用超声远程控制各种蛋白质功能提供了一种可能的途径。
