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Cell Chem Biol. 2019 Oct 17;26(10):1407-1416.e5. doi: 10.1016/j.chembiol.2019.07.007. Epub 2019 Aug 1.

Split-miniSOG for Spatially Detecting Intracellular Protein-Protein Interactions by Correlated Light and Electron Microscopy.

Author information

1
Department of Neurosciences, National Center for Microscopy and Imaging Research, University of California, San Diego, La Jolla, CA 92093, USA. Electronic address: dboassa@ucsd.edu.
2
Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA.
3
Department of Neurosciences, National Center for Microscopy and Imaging Research, University of California, San Diego, La Jolla, CA 92093, USA.
4
Department of Biomedical Engineering and Biological Design Center, Boston University, Boston, MA 02215, USA.
5
Department of Neurosciences, National Center for Microscopy and Imaging Research, University of California, San Diego, La Jolla, CA 92093, USA; Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA.
6
Department of Biomedical Engineering and Biological Design Center, Boston University, Boston, MA 02215, USA. Electronic address: jtngo@bu.edu.

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.

KEYWORDS:

LOV domain; electron microscopy; protein-protein interactions; split-fluorescent proteins

PMID:
31378710
PMCID:
PMC6800643
[Available on 2020-10-17]
DOI:
10.1016/j.chembiol.2019.07.007

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