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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Selective photoinactivation of protein function through environment-sensitive switching of singlet oxygen generation by photosensitizer
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Selective photoinactivation of protein function through environment-sensitive switching of singlet oxygen generation by photosensitizer

机译：通过环境敏感的光敏剂产生单线态氧的转换，选择性地使蛋白质功能发生光失活

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摘要

Chromophore-assisted light inactivation is a promising technique to inactivate selected proteins with high spatial and temporal resolution in living cells, but its use has been limited because of the lack of a methodology to prevent nonspecific photodamage in the cell owing to reactive oxygen species generated by the photosensitizer. Here we present a design strategy for photosensitizers with an environment-sensitive off/on switch for singlet oxygen (~1O_2) generation, which is switched on by binding to the target, to improve the specificity of protein photoinactivation. ~1O_2 generation in the unbound state is quenched by photoinduced electron transfer, whereas ~1O_2 generation can occur in the hydrophobic environment provided by the target protein, after specific binding. Inositol 1,4,5-trisphosphate receptor, which has been suggested to have a hydrophobic pocket around the ligand binding site, was specifically inactivated by an environment-sensitive photosensitizer-conjugated inositol 1,4,5-trisphosphate receptor ligand without ~1O_2 generation in the cytosol of the target cells, despite light illumination, demonstrating the potential of environment-sensitive photosensitizers to allow high-resolution control of generation of reactive oxygen species in the cell.
机译：发色团辅助的光灭活是一种有希望的技术，可以使活细胞中具有高时空分辨率的选定蛋白质失活，但是由于缺乏防止由细胞产生的活性氧而导致的细胞非特异性光损伤的方法，其使用受到了限制。光敏剂。在这里，我们提出了一种光敏剂的设计策略，该光敏剂具有对环境敏感的单线态氧（〜1O_2）生成的关闭/打开开关，该开关通过与靶标结合而打开，以提高蛋白质光灭活的特异性。在未结合状态下，〜1O_2的产生通过光诱导的电子转移而被淬灭，而〜1O_2的产生可以在特异性结合后在靶蛋白提供的疏水环境中发生。肌醇1,4,5-三磷酸受体被认为在配体结合位点周围有一个疏水口袋，被环境敏感的光敏剂偶联的肌醇1,4,5-三磷酸受体配体特异性灭活，没有〜1O_2生成尽管有光照，但靶细胞的胞质溶胶中仍然存在这种物质，证明了对环境敏感的光敏剂具有潜力，可以高分辨率控制细胞中活性氧的产生。

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