Structural basis for reversible photobleaching of a green fluorescent protein homologue
Structural basis for reversible photobleaching of a green fluorescent protein homologue
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DOI：
10.1073/pnas.0700059104
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发表时间：
2007-04-17
影响因子：
11.1
通讯作者：
Remington, S. James
中科院分区：
文献类型：
--
作者：
Henderson, J. Nathan;Ai, Hui-wang;Remington, S. James
荧光蛋白（FP）变体可以在荧光和非荧光状态之间可逆地转换，已被证明是荧光显微镜领域创新的催化剂。然而，对这一进程的结构基础仍然知之甚少。高分辨率结构的FP来自棒形藻在荧光和光诱导的非荧光状态显示，快速和完全的荧光损失后观察到的照明与450 nm的光结果从顺反子异构化的发色团。光诱导的变化，从良好有序的顺式异构体的高度非平面和无序的反式异构体的配置是伴随着一个戏剧性的重排的内部侧链。两者合计，结构提供了一个解释的荧光照射后的损失，缓慢的光无关的恢复，和快速的光诱导的荧光恢复。基本机制似乎是迄今为止报道的所有可光活化和可逆光开关FP所共有的。
Fluorescent protein (FP) variants that can be reversibly converted between fluorescent and nonfluorescent states have proven to be a catalyst for innovation in the field of fluorescence microscopy. However, the structural basis of the process remains poorly understood. High-resolution structures of a FP derived from Clavularia in both the fluorescent and the light-induced nonfluorescent states reveal that the rapid and complete loss of fluorescence observed upon illumination with 450-nm light results from cistrans isomerization of the chromophore. The photoinduced change in configuration from the well ordered cis isomer to the highly nonplanar and disordered trans isomer is accompanied by a dramatic rearrangement of internal side chains. Taken together, the structures provide an explanation for the loss of fluorescence upon illumination, the slow light-independent recovery, and the rapid light-induced recovery of fluorescence. The fundamental mechanism appears to be common to all of the photoactivatable and reversibly photoswitchable FPs reported to date.