Development of a Photoactivatable Proximity Labeling Method for the Identification of Nuclear Proteins
Development of a Photoactivatable Proximity Labeling Method for the Identification of Nuclear Proteins
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DOI：
10.1246/cl.190804
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发表时间：
2020-02-01
影响因子：
1.6
通讯作者：
Hamachi, Itaru
中科院分区：
文献类型：
--
作者：
Tamura, Tomonori;Takato, Mikiko;Hamachi, Itaru
细胞器蛋白质组的精确特征对于在分子水平上破译各种细胞功能是必不可少的。我们在这里报告了一种光活化的邻近标记方法，用于识别具有高时空分辨率的细胞器蛋白质组。在我们的策略中，针对活细胞核的光敏剂被光照射以局部产生单线态氧。然后，一种基于邻苯二胺的底物混杂地标记附近的蛋白质。这种方法与LC-MS/MS联用，可以对核蛋白进行特异性鉴定，包括典型的核蛋白，如组蛋白和层粘连蛋白。
The precise characterization of organellar proteomes is essential for deciphering a variety of cellular functions at the molecular level. We here report a photoactivatable proximity labeling method for identifying organellar proteomes with high spatiotemporal resolution. In our strategy, a photosensitizer targeted to the nucleus of live cells is irradiated with light to locally generate singlet oxygen. An o-phenylenediamine-based substrate then promiscuously tags nearby proteins. This method coupled with LC-MS/MS allowed the specific identification of nuclear proteins, including typical nuclear proteins such as histones and lamins.