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Light-Inducible Generation of Membrane Curvature in Live Cells with Engineered BAR Domain Proteins.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-04-07 , DOI: 10.1021/acssynbio.9b00516
Taylor Jones 1 , Aofei Liu 1 , Bianxiao Cui 1
Affiliation  

Nanoscale membrane curvature is now understood to play an active role in essential cellular processes such as endocytosis, exocytosis, and actin dynamics. Previous studies have shown that membrane curvature can directly affect protein function and intracellular signaling. However, few methods are able to precisely manipulate membrane curvature in live cells. Here, we report the development of a new method of generating nanoscale membrane curvature in live cells that is controllable, reversible, and capable of precise spatial and temporal manipulation. For this purpose, we make use of Bin/Amphiphysin/Rvs (BAR) domain proteins, a family of well-studied membrane-remodeling and membrane-sculpting proteins. Specifically, we engineered two optogenetic systems, opto-FBAR and opto-IBAR, that allow light-inducible formation of positive and negative membrane curvature, respectively. Using opto-FBAR, blue light activation results in the formation of tubular membrane invaginations (positive curvature), controllable down to the subcellular level. Using opto-IBAR, blue light illumination results in the formation of membrane protrusions or filopodia (negative curvature). These systems present a novel approach for light-inducible manipulation of nanoscale membrane curvature in live cells.

中文翻译:

具有工程改造的BAR结构域蛋白的活细胞中光诱导的膜曲率生成。

如今,人们已将纳米级膜曲率理解为在基本细胞过程(如胞吞作用,胞吐作用和肌动蛋白动力学)中发挥积极作用。先前的研究表明,膜曲率可以直接影响蛋白质功能和细胞内信号传导。但是,很少有方法能够精确地控制活细胞中的膜曲率。在这里,我们报告了一种新的方法的发展,该方法可在活细胞中产生纳米级的膜曲率,这种方法可控,可逆且能够进行精确的时空操纵。为此,我们利用Bin / Amphiphysin / Rvs(BAR)域蛋白,这是一个经过充分研究的膜重塑和膜雕刻蛋白家族。具体来说,我们设计了两个光遗传系统,即opto-FBAR和opto-IBAR,分别允许光诱导形成正膜曲率和负膜曲率。使用光FBAR,蓝光激活导致可控制到亚细胞水平的管状膜内陷(正曲率)形成。使用光IBAR,蓝光照射会导致形成膜突出或丝状伪足(负曲率)。这些系统为活细胞中的纳米级膜曲率的光诱导操纵提供了一种新颖的方法。
更新日期:2020-04-23
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