Highly Efficient Fluorescent Probe to Visualize Protein Interactions at the Superresolution

ACS Chem Biol. 2024 Jun 21;19(6):1271-1279. doi: 10.1021/acschembio.4c00075. Epub 2024 Jun 4.

Abstract

Superresolution microscopy (SR microscopy) of protein-protein interactions (PPIs) occurring in subcellular structures is essential for understanding cellular functions. However, a powerful and useful technology for SR microscopy of PPIs remains elusive. Here, we develop a highly efficient photoconvertible fluorescent probe, named split-Dendra2, for SR microscopy of PPIs in the cell. We found that split-Dendra2 enables a highly efficient detection of PPIs, making it possible to perform SR microscopy of PPIs with high spatial resolution and high image reconstruction fidelity. We demonstrate the utility of split-Dendra2 by visualizing PPIs occurring in small subcellular structures at the superresolution, such as clathrin-coated pits and focal adhesions, which cannot be visualized by the existing tools. Split-Dendra2 offers a powerful and useful tool that greatly expands the possibility of SR microscopy and can contribute to revealing the function of PPIs at the nanoscale resolution.

MeSH terms

  • Fluorescent Dyes* / chemistry
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence* / methods
  • Protein Interaction Mapping / methods

Substances

  • Fluorescent Dyes