Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China.
Nature. 2019 May;569(7757):509-513. doi: 10.1038/s41586-019-1188-1. Epub 2019 May 8.
A universal gain-of-function approach for selective and temporal control of protein activity in living systems is crucial to understanding dynamic cellular processes. Here we report development of a computationally aided and genetically encoded proximal decaging (hereafter, CAGE-prox) strategy that enables time-resolved activation of a broad range of proteins in living cells and mice. Temporal blockage of protein activity was computationally designed and realized by genetic incorporation of a photo-caged amino acid in proximity to the functional site of the protein, which can be rapidly removed upon decaging, resulting in protein re-activation. We demonstrate the wide applicability of our method on diverse protein families, which enabled orthogonal tuning of cell signalling and immune responses, temporal profiling of proteolytic substrates upon caspase activation as well as the development of protein-based pro-drug therapy. We envision that CAGE-prox will open opportunities for the gain-of-function study of proteins and dynamic biological processes with high precision and temporal resolution.
在活细胞中，一种通用的、可使蛋白质活性具有选择性和时间可控性的功能增益方法对于理解动态细胞过程至关重要。在这里，我们报告了一种计算辅助和基因编码的邻近解笼（简称 CAGE-prox）策略的开发，该策略可在活细胞和小鼠中实现广泛的蛋白质的时间分辨激活。通过在蛋白质的功能位点附近基因整合光笼氨基酸，在计算上设计并实现了蛋白质活性的时间阻断，该氨基酸在解笼后可迅速去除，从而导致蛋白质重新激活。我们证明了我们的方法在多种蛋白质家族中的广泛适用性，这使得细胞信号和免疫反应的正交调谐、半胱天冬酶激活时蛋白酶底物的时间分析以及基于蛋白质的前药治疗成为可能。我们设想 CAGE-prox 将为具有高精度和时间分辨率的蛋白质和动态生物过程的功能增益研究开辟机会。
