Protein coacervates serve as hubs to concentrate and sequester proteins and nucleotides and thus function as membraneless organelles to manipulate cell physiology. We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light. Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light. We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain. Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions. An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae. The methods described here provide novel strategies for inducing protein phase separation using light.

中文翻译：

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分裂：使用光诱导的跃迁进行稳定的蛋白质凝聚。

蛋白质凝聚层充当集中和隔离蛋白质和核苷酸的枢纽，因此起无膜细胞器的作用，以操纵细胞生理。我们已经设计出一种凝聚蛋白来创建可调谐的合成无膜细胞器，这些细胞器会响应单个光脉冲而组装。凝聚由来自蛋白LAF-1的内在无序的RGG域驱动，而光响应性则由蛋白PhoCl编码，该蛋白响应405 nm的光而裂解。我们开发了一种融合蛋白，其中包含一个可溶性麦芽糖结合蛋白结构域，PhoCl和两个RGG结构域副本。在405 nm处照射几秒钟足以裂解PhoCl，除去增溶结构域，并在数分钟内在细胞大小的油包水乳液中实现RGG驱动的凝聚。该系统的优化版本在酿酒酵母中显示出光诱导的凝聚。这里描述的方法提供了使用光诱导蛋白质相分离的新策略。