Highly Efficient Fluorescent Probe to Visualize Protein Interactions at the Superresolution

• Published in: ACS Chemical Biology Vol. 19; no. 6; pp. 1271 - 1279
• Main Authors: Aono, Yuki, Nakajima, Takahiro, Ichimiya, Wataru, Yoshida, Mayumi, Sato, Moritoshi
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 21.06.2024; American Chemical Society (ACS)
• Subjects: Fluorescent Dyes - chemistry; HeLa Cells; Humans; Microscopy, Fluorescence - methods; Protein Interaction Mapping - methods
• ISSN: 1554-8929; 1554-8937; 1554-8937
• DOI: 10.1021/acschembio.4c00075

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.