Chemo-optogenetic protein translocation system using a photoactivatable self-localizing ligand
Chemo-optogenetic protein translocation system using a photoactivatable self-localizing ligand
复制标题
使用光激活自定位配体的化学光遗传学蛋白质易位系统
DOI：
10.1021/acschembio.1c00416
复制
发表时间：
2021
影响因子：
4
通讯作者：
Shinya Tsukiji
中科院分区：
文献类型：
--
作者：
Tatsuyuki Yoshii;Choji Oki;Rei Watahiki;Akinobu Nakamura;Kai Tahara;Keiko Kuwata;Toshiaki Furuta;Shinya Tsukiji
利用光操纵亚细胞蛋白质定位是以高时空精度控制信号过程的一种有效方法。最广泛使用的策略是基于光诱导的蛋白质异二聚化。使用合成的小分子可以控制目标蛋白质的定位，从而在不需要第二种蛋白质的情况下对光做出反应，这有几个优点。然而，这样的方法还没有得到很好的确立。在这里，我们提出了一种使用可光激活的自定位配体(PASL)来控制蛋白质定位的化学-光遗传学方法。我们开发了一种PASL，它可以在光照后几秒钟内将感兴趣的标签融合蛋白从细胞质招募到质膜。这种PASL诱导的蛋白易位(PaSLIPT)是可逆的，能够在活细胞中对信号过程进行时空控制，即使在局部地区也是如此。PaSLIPT还可以用于在单个细胞中实现同时的光刺激和分子过程的多路成像，为询问和工程动态细胞功能提供了一个有吸引力的新颖的化学光遗传平台。
Manipulating subcellular protein localization using light is a powerful approach for controlling signaling processes with high spatiotemporal precision. The most widely used strategy for this is based on light-induced protein heterodimerization. The use of small synthetic molecules that can control the localization of target proteins in response to light without the need for a second protein has several advantages. However, such methods have not been well established. Herein, we present a chemo-optogenetic approach for controlling protein localization using a photoactivatable self-localizing ligand (paSL). We developed a paSL that can recruit tag-fused proteins of interest from the cytoplasm to the plasma membrane within seconds upon light illumination. This paSL-induced protein translocation (paSLIPT) is reversible and enables the spatiotemporal control of signaling processes in living cells, even in a local region. paSLIPT can also be used to implement simultaneous optical stimulation and multiplexed imaging of molecular processes in a single cell, offering an attractive and novel chemo-optogenetic platform for interrogating and engineering dynamic cellular functions.