Reed Ellen H, Schuster Benjamin S, Good Matthew C, Hammer Daniel A
Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
Department of Chemical and Biochemical Engineering, Rutgers University, New Brunswick, New Jersey 08854, United States.
ACS Synth Biol. 2020 Mar 20;9(3):500-507. doi: 10.1021/acssynbio.9b00503. Epub 2020 Feb 28.
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 . The methods described here provide novel strategies for inducing protein phase separation using light.
蛋白质凝聚物作为中心来浓缩和隔离蛋白质及核苷酸，从而作为无膜细胞器发挥作用以调控细胞生理。我们设计了一种凝聚蛋白来创建可调节的合成无膜细胞器，其能响应单次光脉冲进行组装。凝聚由来自蛋白质LAF-1的内在无序RGG结构域驱动，光响应性由蛋白质PhoCl编码，PhoCl会响应405nm光而裂解。我们开发了一种融合蛋白，其包含一个可溶的麦芽糖结合蛋白结构域、PhoCl和两个RGG结构域拷贝。在405nm下照射几秒就足以裂解PhoCl，去除可溶结构域，并在几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……中表现出光诱导凝聚。本文所述方法为利用光诱导蛋白质相分离提供了新策略。
