Atomistic design of microbial opsin-based blue-shifted optogenetics tools.
Atomistic design of microbial opsin-based blue-shifted optogenetics tools.
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DOI：
10.1038/ncomms8177
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发表时间：
2015-05-15
影响因子：
16.6
通讯作者：
Nureki, Osamu
中科院分区：
文献类型：
--
作者：
Kato, Hideaki E.;Kamiya, Motoshi;Sugo, Seiya;Ito, Jumpei;Taniguchi, Reiya;Orito, Ayaka;Hirata, Kunio;Inutsuka, Ayumu;Yamanaka, Akihiro;Maturana, Andres D.;Ishitani, Ryuichiro;Sudo, Yuki;Hayashi, Shigehiko;Nureki, Osamu
具有结合发色团的微生物视蛋白作为光敏离子转运蛋白发挥功能，并且已经在光遗传学中用于神经元活动的光学控制。分子工程已被用于创建颜色变体，用于光遗传学工具的功能增强，但受到蛋白质-发色团相互作用的复杂性的限制。在这里，我们报告的发展蓝移的颜色变体通过合理的设计在原子分辨率，通过精确的混合分子模拟，电生理学和X射线晶体学实现。分子模拟模型和晶体结构揭示了由组合突变引起的发色团精确设计的构象变化，这些突变缩小了其π-共轭系统，再加上静电调谐，使吸收光谱发生最大100 nm的大蓝移，同时保持光敏离子传输活性。我们阐述的设计原则适用于其他微生物视蛋白，并澄清了最近从天然来源分离的微生物视蛋白的蓝移作用光谱的潜在分子机制。 视网膜结合的视蛋白被广泛用于体内神经元活动的光学控制，即所谓的光遗传学。在这里，使用分子模拟，生物化学，电生理学和X射线晶体学，作者提出了新的分子设计原则，用于产生微生物视紫红质的蓝移变体。
Microbial opsins with a bound chromophore function as photosensitive ion transporters and have been employed in optogenetics for the optical control of neuronal activity. Molecular engineering has been utilized to create colour variants for the functional augmentation of optogenetics tools, but was limited by the complexity of the protein–chromophore interactions. Here we report the development of blue-shifted colour variants by rational design at atomic resolution, achieved through accurate hybrid molecular simulations, electrophysiology and X-ray crystallography. The molecular simulation models and the crystal structure reveal the precisely designed conformational changes of the chromophore induced by combinatory mutations that shrink its π-conjugated system which, together with electrostatic tuning, produce large blue shifts of the absorption spectra by maximally 100 nm, while maintaining photosensitive ion transport activities. The design principle we elaborate is applicable to other microbial opsins, and clarifies the underlying molecular mechanism of the blue-shifted action spectra of microbial opsins recently isolated from natural sources. Retinal-bound opsins are widely used tools for optical control of neuronal activity in vivo, so called optogenetics. Here, using molecular simulations, biochemistry, electrophysiology and X-ray crystallography, the authors present new molecular design principles for the generation of blue-shifted variants of microbial rhodopsins.
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影响因子：
4.1
作者：
BENNETT, CH
通讯作者：
BENNETT, CH
影响因子：
62.1
作者：
Ernst, Oliver P.;Lodowski, David T.;Elstner, Marcus;Hegemann, Peter;Brown, Leonid S.;Kandori, Hideki
通讯作者：
Kandori, Hideki
影响因子：
48
作者：
Guo, Zengcai V.;Hart, Anne C.;Ramanathan, Sharad
通讯作者：
Ramanathan, Sharad
影响因子：
5.5
作者：
Fujimoto, Kazuhiro;Hayashi, Shigehiko;Nakatsuji, Hiroshi
通讯作者：
Nakatsuji, Hiroshi
影响因子：
15
作者：
Ferré, N;Olivucci, M
通讯作者：
Olivucci, M