In Situ Formation of an Azo Bridge on Proteins Controllable by Visible Light.
In Situ Formation of an Azo Bridge on Proteins Controllable by Visible Light.
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DOI：
10.1021/jacs.5b06234
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发表时间：
2015-09-09
影响因子：
15
通讯作者：
Wang L
中科院分区：
文献类型：
--
作者：
Hoppmann C;Maslennikov I;Choe S;Wang L
蛋白质的光学调制为理解生物过程提供了优越的时空分辨率，而建立在光敏蛋白质上的光开关已经显著地推动了神经元和细胞的研究。小分子光开关可以通过减少潜在的干扰和提供高度特异性的调制位点来补充基于蛋白质的开关。然而，基因可编码性和对非紫外光的响应性是蛋白质光开关所具有的两个理想特性，但将其设计成小分子光开关是具有挑战性的。在这里，我们开发了一种小分子光开关，可以通过基因原位安装到蛋白质上，并通过可见光控制。设计了一种基于五氟偶氮苯的光开关点击氨基酸(F-PSCaa)对可见光的异构化反应。F-PSCaa通过扩展遗传密码与蛋白质中的基因整合后，与蛋白质中附近的半胱氨酸发生反应，在原位产生偶氮桥。由F-PSCaa构建的桥可以在可见光下切换，并允许在这种光下调节CaM的构象和结合。这种光开关因其最小的干扰、位置的灵活性、遗传可编码性和对更生物兼容的可见光的响应而在光生物学中被证明是有价值的。
Optical modulation of proteins provides superior spatiotemporal resolution for understanding biological processes, and photoswitches built on light-sensitive proteins have been significantly advancing neuronal and cellular studies. Small molecule photoswitches could complement protein-based switches by mitigating potential interference and affording high specificity for modulation sites. However, genetic encodability and responsiveness to non-ultraviolet light, two desired properties possessed by protein photoswitches, are challenging to be engineered into small molecule photoswitches. Here we developed a small molecule photoswitch that can be genetically installed onto proteins in situ and controlled by visible light. A pentafluoro azobenzene based photoswitchable click amino acid (F-PSCaa) was designed to isomerize in response to visible light. After genetic incorporation into proteins via the expansion of the genetic code, F-PSCaa reacts with a nearby cysteine within the protein generating an azo bridge in situ. The resultant bridge built by F-PSCaa is switchable by visible light and allows conformation and binding of CaM to be regulated by such light. This photoswitch should prove valuable in optobiology for its minimal interference, site flexibility, genetic encodability, and response to the more biocompatible visible light.