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Time-resolved protein activation by proximal decaging in living systems

机译：生命系统中近端递减的时间分辨蛋白质活化

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摘要

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.
机译：在生命系统中选择性和暂时控制蛋白质活性的通用功能获得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遗传编码的近端递减（以下称为CAGE-prox）策略的开发，该策略能够在时间上激活活细胞和小鼠中广泛的蛋白质。蛋白质活性的暂时性阻滞是通过在蛋白质的功能部位附近遗传引入光笼氨基酸而设计和实现的，该氨基酸可以在降解后迅速去除，从而导致蛋白质重新活化。我们证明了我们的方法在各种蛋白质家族中的广泛适用性，这使得细胞信号传导和免疫应答的正交调节，胱天蛋白酶激活后蛋白水解底物的时间分布以及基于蛋白质的药物治疗的发展成为可能。我们设想，CAGE-prox将为蛋白质和动态生物过程的高精度和时间分辨率的功能获得研究提供机会。

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  • 来源
    《Nature》 |2019年第7757期|509-513|共5页
  • 作者

    Wang Jie; Liu Yuan; Liu Yanjun; Zheng Siqi; Wang Xin; Zhao Jingyi; Yang Fan; Zhang Gong; Wang Chu; Chen Peng R.;

  • 作者单位

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China|Peking Univ, Acad Adv Interdisciplinary Studies, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China|Peking Univ, Acad Adv Interdisciplinary Studies, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China|Peking Univ, Acad Adv Interdisciplinary Studies, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China|Peking Univ, Minist Educ, Key Lab Bioorgan Chem & Mol Engn, Beijing, Peoples R China|Peking Univ, Peking Tsinghua Ctr Life Sci, Beijing, Peoples R China;

    Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Synthet & Funct Biomol Ctr, Beijing, Peoples R China|Peking Univ, Acad Adv Interdisciplinary Studies, Beijing, Peoples R China|Peking Univ, Minist Educ, Key Lab Bioorgan Chem & Mol Engn, Beijing, Peoples R China|Peking Univ, Peking Tsinghua Ctr Life Sci, Beijing, Peoples R China;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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