- Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy.
Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy.
Cell chemical biology (2019-08-06)
Daniela Boassa, Sakina P Lemieux, Varda Lev-Ram, Junru Hu, Qing Xiong, Sebastien Phan, Mason Mackey, Ranjan Ramachandra, Ryan Emily Peace, Stephen R Adams, Mark H Ellisman, John T Ngo
PMID31378710
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.
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