Homo-molecular Fluorescence Complementation for Direct Visualization of Receptor Oligomerization in Living Cells

Angew Chem Int Ed Engl. 2019 Feb 11;58(7):2045-2049. doi: 10.1002/anie.201812780. Epub 2019 Jan 14.

Abstract

Cell surface receptor oligomerization is an attractive target process for drug screening. However, simple but reliable (and thus high-throughput) visualization methods for receptor oligomerization are still lacking. Herein, we report on a new single-construct homo-molecular fluorescence complementation (Homo-FC) probe, which shows strong fluorescence signals by oligomerization of fused receptors in living cells with unexpectedly low background signals. Importantly, this high signal-to-noise ratio was not affected by expression level variations of fused receptors. The Homo-FC probe was developed by optimized flopped fusion of split fragments of superfolder green fluorescence protein and subsequent surface charge engineering. Homo-FC reliably visualized the oligomerization of diverse natural receptors such as GPCR, EGFR, and even cytosolic DAI.

Keywords: drug discovery; fluorescence complementation; imaging; membrane receptor oligomerization; protein engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescence*
  • Fluorescent Dyes / chemistry*
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence
  • Models, Molecular
  • Optical Imaging*
  • Receptors, Cell Surface / analysis*

Substances

  • Fluorescent Dyes
  • Receptors, Cell Surface