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首页> 外文期刊>Journal of Molecular Biology >Optogenetic Navigation of Routes Leading to Protein Amyloidogenesis in Bacteria
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Optogenetic Navigation of Routes Leading to Protein Amyloidogenesis in Bacteria

机译：导致蛋白质淀粉膜过生的途径的光学导航

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Modulation of liquid-liquid and liquid-hydrogel phase transitions is central to avoid the cytotoxic aggregation of proteins in eukaryotic cells, but knowledge on its relevance in bacteria is limited. Here the power of optogenetics to engineer proteins as light-responsive switches has been used to control the balance between solubility and aggregation for LOV2-WH1, a chimera between the plant blue light-responsive domain LOV2 and the bacterial prion-like protein RepA-WH1. These proteins were first linked by fusing, as a continuous alpha-helix, the C-terminal photo-transducer J alpha helix in LOV2 with the N-terminal domain-closure alpha 1 helix in RepA-WH1, and then improved for light-responsiveness by including mutations in the J alpha moiety. In the darkness and in a crowded solution in vitro, LOV2-WH1 nucleates the irreversible assembly of amyloid fibers into a hydrogel. However, under blue light illumination, LOV2-WH1 assembles as soluble oligomers. When expressed in Escherichia coli, LOV2-WH1 forms in the darkness large intracellular amyloid inclusions compatible with bacterial proliferation. Strikingly, under blue light, LOV2-WH1 aggregates decrease in size, while they become detrimental for bacterial growth. LOV2-WH1 optogenetics governs the assembly of mutually exclusive inert amyloid fibers or cytotoxic oligomers, thus enabling the navigation of the conformational landscape.of protein amyloidogenesis to generate potential photo-activated anti-bacterial devices (optobiotics). (C) 2019 Elsevier Ltd. All rights reserved.
机译：液体和液态水凝胶相转变的调节是避免真核细胞中蛋白质细胞毒性聚集的中央，但是关于其在细菌中的相关知识有限。这里，光源对工程蛋白作为光响应开关的功率已经用于控制LOV2-WH1的溶解度和聚集之间的平衡，植物蓝光响应域LOV2和细菌朊病毒蛋白质REPA-WH1之间的嵌合性。这些蛋白质首先通过熔化，作为连续α-螺旋，在LOV2中的C末端光旋转器Jα螺旋中与REBA-WH1中的N末端域闭合α1螺旋一起连接，然后改善光响应性通过包括Jα部分中的突变。在黑暗中和在一个拥挤的解决方案中，Lov2-Wh1核成淀粉样纤维的不可逆组装到水凝胶中。然而，在蓝光照明下，LOV2-WH1组装为可溶性低聚物。当在大肠杆菌中表达时，LOV2-WH1在黑暗中形成，与细菌增殖相容的大的细胞内淀粉样蛋白含。在蓝光下令人惊讶的是，Lov2-Wh1聚集在尺寸下降，同时对细菌生长有害。 Lov2-WH1 Optogenetics治理相互排斥的惰性淀粉样纤维或细胞毒性低聚物的组装，从而能够导航蛋白质淀粉化物的构象景观。 （c）2019 Elsevier Ltd.保留所有权利。

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