Computational design of chemogenetic and optogenetic split proteins
Computational design of chemogenetic and optogenetic split proteins
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DOI：
10.1038/s41467-018-06531-4
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发表时间：
2018-10-02
影响因子：
16.6
通讯作者：
Dokholyan, Nikolay, V
中科院分区：
文献类型：
--
作者：
Dagliyan, Onur;Krokhotin, Andrey;Dokholyan, Nikolay, V
用化学遗传学和光遗传学控制蛋白质活性已被证明对于测试关于快速生物过程中蛋白质功能的假设是强大的。通过分裂蛋白质然后通过诱导重组来拯救它们的活性来控制蛋白质，这为控制不同的蛋白质活性提供了巨大的潜力。由于自发组装、难以识别适当的分裂位点以及有效重组的低效诱导，构建分裂蛋白一直是困难的。在这里，我们提出了一种自动化的方法来设计有效的分裂蛋白调节配体或光（SPELL）。我们开发了一个评分功能与工程结构域，使重组蛋白质的一半，具有高效率和减少自发组装。我们通过将其应用于活细胞中各种形状和大小的蛋白质来证明SPELL。SPELL服务器（spell.dokhlab.org）提供了分裂站点的自动预测。
Controlling protein activity with chemogenetics and optogenetics has proven to be powerful for testing hypotheses regarding protein function in rapid biological processes. Controlling proteins by splitting them and then rescuing their activity through inducible reassembly offers great potential to control diverse protein activities. Building split proteins has been difficult due to spontaneous assembly, difficulty in identifying appropriate split sites, and inefficient induction of effective reassembly. Here we present an automated approach to design effective split proteins regulated by a ligand or by light (SPELL). We develop a scoring function together with an engineered domain to enable reassembly of protein halves with high efficiency and with reduced spontaneous assembly. We demonstrate SPELL by applying it to proteins of various shapes and sizes in living cells. The SPELL server (spell.dokhlab.org ) offers an automated prediction of split sites.