黄培元, 宋禹桐, 张宁, 赵志豪, 段丽婷. 基于光控蛋白质相互作用的光遗传学技术及其应用. 中国激光, 2020, 47(2): 0207010。 Peiyuan Huang, Yutong Song, Ning Zhang, Zhihao Zhao, Liting Duan..Optogenetics Based on Light-Gated Protein-Protein Interactions and Its Applications. Chinese Journal of Lasers, 2020, 47(2): 0207010.
Department of Biomedical Engineering, The Chinese University of Hong Kong, Hong Kong 123456, China
West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, Sichuan 610041, China;
Optogenetics is an emerging technique that exploits light to control cells by combining optics and genetics techniques. In optogenetic systems, cells are genetically modified to express photosensitive proteins and consequently become responsive to light pulses. Optogenetics has revolutionized neuroscience research by facilitating selective and rapid control of targeted neurons expressing light-gated ion channels. In addition to light-gated ion channels, photosensitive proteins based on light-gated protein-protein interactions are widely used in optogenetic research. In this review, we discuss these common photosensitive proteins and summarize optogenetic applications in optical control of gene expression, phase separation, biosynthesis, and organelle distribution based on light-gated protein-protein interactions.
Key words medical optics; optogenetics; protein-protein interaction; gene expression; organelle transport; phase separation; biosynthesis
光敏蛋白是指接受特定波长的光刺激之后能够改变自身结构的一类蛋白质分子。光敏蛋白存在于自然界多种生物体中,使生物能够对环境光照变化做出反应。有些光敏蛋白接受特定波长的光刺激后,会与另一个蛋白发生异源二聚化(hetero-dimerization)[图1(a)],而有些光敏蛋白会发生同源二聚化或多聚化(homo-dimerization or homo-oligomerization)[图1(b)]。这两类光调节的蛋白-蛋白相互作用被广泛使用于光遗传技术当中以实现光控各种细胞行为。本研究主要介绍三类广泛使用的光敏蛋白,分别是光敏色素(phytochrome)、隐花色素(cryptochrome)和光-氧-电感应结构域(light-oxygen-voltage-sensing domain)的性质、特点和功能。
Two types of light-gated protein-protein interaction widely used in optogenetics. (a) Hetero-dimerization; (b) homo-dimerization
Schematic diagram of the Corelets system. Corelets system consists of two modules: a GFP-tagged ferritin core functionalized by 24 photoactivatable iLID domains and iLID's cognate partner, sspB, mCherry-tagged and conjugated to IDR. Dashed lines designate light-inducible hetero-dimerizing units
图4Corelets系统示意图。 Corelets 系统包括两个模块:第一个是GFP标记的24个可光激活的iLID连接组成的铁蛋白核心; 第二个是带有mCherry的结合蛋白SspB,并与固有无序蛋白区域IDR缀合。 虚线表示光诱导异源二聚体单元Fig. 4Schematic diagram of the Corelets system. Corelets system consists of two modules: a GFP-tagged ferritin core functionalized by 24 photoactivatable iLID domains and iLID's cognate partner, sspB, mCherry-tagged and conjugated to IDR. Dashed lines designate light-inducible hetero-dimerizing units
图5光触发活性酶蛋白簇聚集。代谢通路导向主要产物,副产物的生成被抑制Fig. 5Gather of light-induced active enzyme protein clusters, in which the input is shunted toward major product, and the production of unwanted byproduct is suppressed
图6使用光遗传学工具控制细胞器运输与分子马达行为的基本机制Fig. 6Basic mechanism optogenetic control of organelle transport and molecular motors
大多数已有的光遗传工具都可被用于这个可逆的、作用于特定范围的控制体系的构建。Duan等把CRY2连接在线粒体、过氧化物酶体或溶酶体上;CIB1则与KIF5A(Kinesin heavy chain isoform 5A,驱动蛋白重链5A亚型)或与BICDN(N terminus of Protein bicaudal D,动力蛋白双锥蛋白D的N端)相融合。在蓝光信号的刺激下,CRY2与CIB1发生异源二聚化,分子马达被召集至细胞器,并开始引导细胞器向微管正端(细胞边缘方向)或负端(细胞核方向)运动。该方法可实现空间上的控制:只有受到蓝光照射的范围内的细胞器发生了明显的分布变化。同时,该方法是可逆的,停止光照后,线粒体在3 h内逐渐恢复了蓝光激发前的分布位置。van Bergeijk等使用LOV结构域二聚体,在不影响其他细胞器的分布的情况下控制了过氧化物酶体在细胞内的定向运输,并展示了在海马体神经元细胞中光控Rab-11(Ras-related protein Rab-11A)的运输和定位可以增强或抑制轴突生长锥生长这一现象。Harterink等把这个系统应用在秀丽隐杆线虫(caenorhabditis elegans)中实现了各种细胞器的运输。这些研究证明了使用光遗传学工具在活体内控制细胞器运输的可行性。
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