Orthogonal Optical Control of a G Protein-Coupled Receptor with a SNAP-Tethered Photochromic Ligand.
Orthogonal Optical Control of a G Protein-Coupled Receptor with a SNAP-Tethered Photochromic Ligand.
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DOI：
10.1021/acscentsci.5b00260
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发表时间：
2015-10-28
影响因子：
18.2
通讯作者：
Trauner D
中科院分区：
文献类型：
--
作者：
Broichhagen J;Damijonaitis A;Levitz J;Sokol KR;Leippe P;Konrad D;Isacoff EY;Trauner D
人工合成光开关的共价连接是一种将光敏感度传递给天然受体的一般方法。它模仿了自然光感受器的逻辑，极大地扩展了光遗传学的研究范围。在这里，我们描述了一种新颖的光开关设计--可光开关的正交远程拴系配体(PORTL)--它将遗传编码的SNAP标签与光致变色配体通过长而灵活的连接物连接到苄基鸟嘌呤上。我们使用这种方法将G蛋白偶联受体mGluR2，一种代谢性谷氨酸受体，转化为光受体(SNAG-mGluR2)，从而对mGluR2的神经元功能提供有效的光学控制：突触前抑制和兴奋性控制。PORTL方法能够使用不同的生物结合方法对不同的天然受体进行多路光学控制。由于SNAP标记已经被证明是可靠的，许多标记SNAP的受体已经可用，而且长链上的光致变色配体很容易设计和合成，因此它应该是广泛适用的。SNAP标记的代谢型谷氨酸受体通过远程拴系的偶氮苯谷氨酸光开关被赋予光敏感性，并可用于用光可逆地诱导GPCR激活来控制神经功能。
The covalent attachment of synthetic photoswitches is a general approach to impart light sensitivity onto native receptors. It mimics the logic of natural photoreceptors and significantly expands the reach of optogenetics. Here we describe a novel photoswitch design—the photoswitchable orthogonal remotely tethered ligand (PORTL)—that combines the genetically encoded SNAP-tag with photochromic ligands connected to a benzylguanine via a long flexible linker. We use the method to convert the G protein-coupled receptor mGluR2, a metabotropic glutamate receptor, into a photoreceptor (SNAG-mGluR2) that provides efficient optical control over the neuronal functions of mGluR2: presynaptic inhibition and control of excitability. The PORTL approach enables multiplexed optical control of different native receptors using distinct bioconjugation methods. It should be broadly applicable since SNAP-tags have proven to be reliable, many SNAP-tagged receptors are already available, and photochromic ligands on a long leash are readily designed and synthesized. A SNAP-tagged metabotropic glutamate receptor is endowed with light sensitivity by using a remotely tethered azobenzene glutamate photoswitch and can be used to induce GPCR activation reversibly with light to control neural function.