Open AccessDevelopment of bimolecular fluorescence complementation using rsEGFP2 for detection and super-resolution imaging of protein-protein interactions in live cells
Author(s) -
Sheng Wang,
Miao Ding,
Xuanze Chen,
Lei Chao,
Yujie Sun
Publication year - 2017
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.8.003119
Subject(s) - bimolecular fluorescence complementation , live cell imaging , fluorescence lifetime imaging microscopy , fluorescent protein , fluorescence microscope , microscopy , fluorescence , resolution (logic) , protein–protein interaction , temporal resolution , sted microscopy , superresolution , biophysics , nanotechnology , green fluorescent protein , chemistry , microbiology and biotechnology , biology , materials science , cell , physics , computer science , optics , biochemistry , artificial intelligence , gene , image (mathematics)
Direct visualization of protein-protein interactions (PPIs) at high spatial and temporal resolution in live cells is crucial for understanding the intricate and dynamic behaviors of signaling protein complexes. Recently, bimolecular fluorescence complementation (BiFC) assays have been combined with super-resolution imaging techniques including PALM and SOFI to visualize PPIs at the nanometer spatial resolution. RESOLFT nanoscopy has been proven as a powerful live-cell super-resolution imaging technique. With regard to the detection and visualization of PPIs in live cells with high temporal and spatial resolution, here we developed a BiFC assay using split rsEGFP2, a highly photostable and reversibly photoswitchable fluorescent protein previously developed for RESOLFT nanoscopy. Combined with parallelized RESOLFT microscopy, we demonstrated the high spatiotemporal resolving capability of a rsEGFP2-based BiFC assay by detecting and visualizing specifically the heterodimerization interactions between Bcl-x L and Bak as well as the dynamics of the complex on mitochondria membrane in live cells.