Split-miniSOG 用于通过相关光和电子显微镜术空间检测细胞内蛋白质-蛋白质相互作用。
Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy.
机构信息
Department of Neurosciences, National Center for Microscopy and Imaging Research, University of California, San Diego, La Jolla, CA 92093, USA.
Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA.
出版信息
Cell Chem Biol. 2019 Oct 17;26(10):1407-1416.e5. doi: 10.1016/j.chembiol.2019.07.007. Epub 2019 Aug 1.
Abstract
A protein-fragment complementation assay (PCA) for detecting and localizing intracellular protein-protein interactions (PPIs) was built by bisection of miniSOG, a fluorescent flavoprotein derived from the light, oxygen, voltage (LOV)-2 domain of Arabidopsis phototropin. When brought together by interacting proteins, the fragments reconstitute a functional reporter that permits tagged protein complexes to be visualized by fluorescence light microscopy (LM), and then by standard as well as "multicolor" electron microscopy (EM) via the photooxidation of 3-3'-diaminobenzidine and its derivatives.
摘要
一种用于检测和定位细胞内蛋白质-蛋白质相互作用（PPIs）的蛋白片段互补测定（PCA），是通过将来源于拟南芥光受体光敏素的 LOV-2 结构域的荧光黄素蛋白 miniSOG 进行二分法构建的。当通过相互作用的蛋白质聚集在一起时，这些片段重新构成一个功能报告器，使得标记的蛋白质复合物能够通过荧光显微镜（LM）可视化，然后通过标准和“多色”电子显微镜（EM）通过 3-3'-二氨基联苯胺及其衍生物的光氧化来实现。
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