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Time-resolved protein activation by proximal decaging in living systems

机译：在生活系统中近端辨中的时间分辨蛋白激活

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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：用于在生活系统中对蛋白质活性的选择性和时间控制的普遍函数和时间控制对于理解动态细胞过程至关重要。在这里，我们报告开发计算辅助和遗传编码的近端码（以下，笼子 - Prox）策略，其能够在活细胞和小鼠中进行多种蛋白质的时间分辨激活。通过遗传掺入邻近蛋白质的功能部位的遗传掺入光笼氨基酸的遗传掺入蛋白质活性的时间堵塞，可以在沉重时迅速除去，导致蛋白质再激活。我们展示了我们对多种蛋白质家族的方法的广泛适用性，该方法能够使细胞信号传导和免疫应答的正交调整，蛋白质溶液在胱天冬酶活动下的时间谱分析以及基于蛋白质的亲药物治疗的发育。我们设想笼式PROX将开放蛋白质和动态生物过程的功能研究的机会，具有高精度和时间分辨率。

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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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