Spatiotemporal control of cell signalling using a light-switchable protein interaction.
Spatiotemporal control of cell signalling using a light-switchable protein interaction.
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作者：
可遗传编码的光学报告分子，如绿色荧光蛋白，已经彻底改变了细胞状态的观察和测量。然而，使用光来精确控制细胞行为的反向挑战直到最近才开始得到解决;半合成的发色团束缚受体和天然存在的通道视紫红质已被用于直接干扰神经元网络。工程化光敏蛋白的困难仍然是大多数细胞生物过程的光学控制的重大障碍。在这里，我们展示了使用一种新的遗传编码的光控制系统的基础上优化可逆的蛋白质-蛋白质相互作用的光敏色素信号网络的拟南芥。因为蛋白质-蛋白质相互作用是细胞信息的最普遍的货币之一，所以这个系统原则上可以被普遍地用于控制各种功能。在这里，我们表明，该系统可以用于精确和可逆地将目标蛋白质转位到膜上，具有微米级的空间分辨率和秒级的时间分辨率。我们表明，光门控易位的上游激活剂的rho家族GTPases，控制肌动蛋白细胞骨架，可用于精确地重塑和指导哺乳动物细胞的细胞形态。在这项工作中优化的光门控蛋白质-蛋白质相互作用应该有助于设计不同的光可编程试剂，可能使新一代的微扰，定量实验在细胞生物学。
Genetically-encodable optical reporters, such as Green Fluorescent Protein, have revolutionized the observation and measurement of cellular states. However, the inverse challenge of using light to precisely control cellular behavior has only recently begun to be addressed; semi-synthetic chromophore-tethered receptors and naturally-occurring channel rhodopsins have been used to directly perturb neuronal networks. The difficulty of engineering light sensitive proteins remains a significant impediment to the optical control to most cell-biological processes. Here we demonstrate the use of a new genetically-encoded light-control system based on an optimized reversible protein-protein interaction from the phytochrome signaling network of Arabidopsis thaliana. Because protein-protein interactions are one of the most general currencies of cellular information, this system can in principal be generically used to control diverse functions. Here we show that this system can be used to precisely and reversibly translocate target proteins to the membrane with micrometer spatial resolution and second time resolution. We show that light-gated translocation of the upstream activators of rho-family GTPases, which control the actin cytoskeleton, can be used to precisely reshape and direct the cell morphology of mammalian cells. The light-gated protein-protein interaction that has been optimized in this work should be useful for the design of diverse light-programmable reagents, potentially enabling a new generation of perturbative, quantitative experiments in cell biology.
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影响因子：
4.5
作者：
Genoud, Thierry;Schweizer, Fabian;Tscheuschler, Anke;Debrieux, Dimitry;Casal, Jorge J.;Schaefer, Eberhard;Hiltbrunner, Andreas;Fankhauser, Christian
通讯作者：
Fankhauser, Christian
DOI：
10.1126/science.1159052
发表时间：
2008-10-17
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Lee J;Natarajan M;Nashine VC;Socolich M;Vo T;Russ WP;Benkovic SJ;Ranganathan R
通讯作者：
Ranganathan R
影响因子：
56.9
作者：
Mettetal, Jerome T.;Muzzey, Dale;van Oudenaarden, Alexander
通讯作者：
van Oudenaarden, Alexander
影响因子：
25
作者：
Boyden, ES;Zhang, F;Deisseroth, K
通讯作者：
Deisseroth, K
影响因子：
64.8
作者：
Ni, M;Tepperman, JM;Quail, PH
通讯作者：
Quail, PH