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

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.

SUBMITTER: Boassa D 

PROVIDER: S-EPMC6800643 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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

Boassa Daniela D   Lemieux Sakina P SP   Lev-Ram Varda V   Hu Junru J   Xiong Qing Q   Phan Sebastien S   Mackey Mason M   Ramachandra Ranjan R   Peace Ryan Emily RE   Adams Stephen R SR   Ellisman Mark H MH   Ngo John T JT  

Cell chemical biology 20190801 10

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 "multic  ...[more]