Optical Control of Phosphoinositide Binding: Rapid Activation of Subcellular Protein Translocation and Cell Signaling.
Optical Control of Phosphoinositide Binding: Rapid Activation of Subcellular Protein Translocation and Cell Signaling.
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DOI：
10.1021/acssynbio.1c00328
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发表时间：
2021-11-19
影响因子：
4.7
通讯作者：
Deiters A
中科院分区：
文献类型：
--
作者：
Ryan A;Hammond GRV;Deiters A
细胞利用蛋白质易位到特定区室来调节蛋白质活性的空间和时间，特别是在信号传导过程中。蛋白质识别和与各种亚细胞膜的结合是由磷脂酰肌醇磷酸 (PIP) 物质网络介导的，这些物质在极性肌醇头上带有一个或多个磷酸部分。在这里，我们报告了一种新的高效方法，通过位点特异性掺入光笼氨基酸来光学控制蛋白质定位，以空间和静电破坏肌醇磷酸盐识别和结合。我们通过光笼罩两种不相关的蛋白质、分选 nexin 3 (SNX3) 和磷脂酶 C delta 1 (PLCδ1) 的普莱克斯特林同源 (PH) 结构域，以及两个不同的 PIP 结合结构域和不同的亚细胞定位，证明了该方法的普遍适用性。我们通过对 Son of Sevenless 2 (SOS2) 的应用，确定了该方法的适用性，SOS2 是一种参与细胞外信号调节激酶/丝裂原激活蛋白激酶 (ERK/MAPK) 级联的信号蛋白。将光笼质膜靶向构建体 PH 增强的绿色荧光蛋白 (EGFP) 与 SOS2 的催化结构域融合后，我们证明了该构建体的光诱导膜定位，导致 NIH 3T3 细胞中 ERK 信号通路快速而广泛的激活。这种方法可以很容易地扩展到其他蛋白质，只需最少的蛋白质工程，并提供一种快速、完全激活的蛋白质易位的急性光学控制方法。
Cells utilize protein translocation to specific compartments for spatial and temporal regulation of protein activity, in particular in the context of signaling processes. Protein recognition and binding to various subcellular membranes is mediated by a network of phosphatidylinositol phosphate (PIP) species bearing one or multiple phosphate moieties on the polar inositol head. Here, we report a new, highly efficient method for optical control of protein localization through the site-specific incorporation of a photocaged amino acid for steric and electrostatic disruption of inositol phosphate recognition and binding. We demonstrate general applicability of the approach by photocaging two unrelated proteins, sorting nexin 3 (SNX3) and the pleckstrin homology (PH) domain of phospholipase C delta 1 (PLCδ1), with two distinct PIP binding domains and distinct subcellular localizations. We have established the applicability of this methodology through its application to Son of Sevenless 2 (SOS2), a signaling protein involved in the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) cascade. Upon fusing the photocaged plasma membrane-targeted construct PH-enhanced green fluorescent protein (EGFP), to the catalytic domain of SOS2, we demonstrated light-induced membrane localization of the construct resulting in fast and extensive activation of the ERK signaling pathway in NIH 3T3 cells. This approach can be readily extended to other proteins, with minimal protein engineering, and provides a method for acute optical control of protein translocation with rapid and complete activation.
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影响因子：
7.8
作者：
Courtney T;Deiters A
通讯作者：
Deiters A
使用光塑料 - 可透明的化学二聚体对活细胞中蛋白质定位的可逆控制。
影响因子：
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作者：
Aonbangkhen C;Zhang H;Wu DZ;Lampson MA;Chenoweth DM
通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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影响因子：
16.6
作者：
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通讯作者：
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