Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy. Issue 10 (17th October 2019)

• Record Type: Journal Article
• Authors: Boassa, Daniela; Lemieux, Sakina P.; Lev-Ram, Varda; Hu, Junru; Xiong, Qing; Phan, Sebastien; Mackey, Mason; Ramachandra, Ranjan; Peace, Ryan Emily; Adams, Stephen R.; Ellisman, Mark H.; Ngo, John T.
• Is Part Of: Cell chemical biology. Volume 26:Issue 10(2019)
• Page Start: 1407
• Page End: 1416.e5
• Publication Date: 2019-10-17
• Subjects: electron microscopy -- protein-protein interactions -- LOV domain -- split-fluorescent proteins; Biochemistry -- Periodicals; 572.05
• DOI: 10.1016/j.chembiol.2019.07.007
• Languages: English
• ISSNs: 2451-9456

Abstract

Summary: 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. Graphical Abstract: Highlights: Split-miniSOG allows intracellular protein-protein interactions to be visualized by correlated fluorescence and electron microscopy Split-miniSOG complementation is reversible Reconstituted complexes are imaged by fluorescence microscopy and TEM/multicolor EM Abstract : Boassa et al. describe the use of split-miniSOG for the visualization of protein aggregates associated with neurodegenerative diseases. This study shows the general utility of this reversible system for detection of spatial organization of molecular complexes in mammalian cells at nanometer resolution.