Structural basis for the phototoxicity of the fluorescent protein KillerRed
Structural basis for the phototoxicity of the fluorescent protein KillerRed
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DOI：
10.1016/j.febslet.2009.07.041
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发表时间：
2009-09-03
期刊：
影响因子：
3.5
通讯作者：
Bourgeois, Dominique
中科院分区：
文献类型：
--
作者：
Carpentier, Philippe;Violot, Sebastien;Bourgeois, Dominique
据报道，红色荧光蛋白 KillerRed 由水螅色素蛋白 anm2CP 改造而来，可通过发色团辅助光灭活 (CALI) 效应诱导强烈的细胞毒性。在这里，我们展示了 KillerRed 天然状态和漂白状态的 X 射线结构。显露出一条长长的充满水的通道，将发色团的亚甲基桥连接到溶剂。该通道促进氧气和与激发的发色团反应形成的活性氧 (ROS) 的转运。讨论了用于产生 KillerRed 的关键突变的功能作用，揭示了漂白状态下的强烈发色团扭曲，并提出了 ROS 产生和自我保护的机制。其特征在于存在部分成熟、耐光、发绿光的状态，这解释了“预漂白”KillerRed 增强 CALI 的原因。 (C) 2009 年欧洲生化学会联合会。由 Elsevier B.V 出版。保留所有权利。
The red fluorescent protein KillerRed, engineered from the hydrozoan chromoprotein anm2CP, has been reported to induce strong cytotoxicity through the chromophore assisted light inactivation (CALI) effect. Here, we present the X-ray structures of KillerRed in its native and bleached states. A long water-filled channel is revealed, connecting the methylene bridge of the chromophore to the solvent. This channel facilitates the transit of oxygen and of reactive oxygen species (ROS) formed by reaction with the excited chromophore. The functional roles of key mutations used to produce KillerRed are discussed, strong chromophore distortions in the bleached state are revealed, and mechanisms for ROS production and self protection are proposed. The presence of a partially mature, photo-resistant, green-emitting state is characterized, which accounts for enhanced CALI by "pre-bleached" KillerRed. (C) 2009 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.