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Updated: Jul 12, 2025

Imaging Protein-protein Interactions in vivo

Imaging Protein-protein Interactions in vivo

Published on: October 10, 2010

Imaging Proteins Sensitive to Direct Fusions Using Transient Peptide-Peptide Interactions.

Zoe Gidden^1,2, Curran Oi³, Emily J Johnston^1,4

¹School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3DW, U.K.

|November 2, 2023

Summary

This summary is machine-generated.

This study introduces LIVE-PAINT, a super-resolution microscopy technique using short peptides to visualize proteins in live cells. It successfully images yeast membrane proteins and allows for concurrent nanoscale imaging of multiple proteins.

Keywords:

LIVE-PAINT live-cell imaging membrane protein protein−protein interaction super-resolution microscopy yeast

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Last Updated: Jul 12, 2025

Imaging Protein-protein Interactions in vivo

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Published on: October 10, 2010

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Bimolecular Fluorescence Complementation

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Published on: April 15, 2011

Area of Science:

Cell biology
Microscopy
Biophysics

Background:

Fluorescence microscopy is crucial for visualizing protein location and function in living cells.
Direct fusion of fluorescent proteins can alter protein behavior, hindering live-cell studies.
Standard fluorescence microscopy resolution is insufficient to resolve individual proteins.

Purpose of the Study:

To overcome limitations of direct fluorescent protein fusions in live-cell protein studies.
To apply and validate the LIVE-PAINT super-resolution technique for challenging protein targets.
To demonstrate the capability of LIVE-PAINT for multi-protein nanoscale imaging.

Main Methods:

Development and application of LIVE-PAINT (Localization by Interaction of Viral-like Entities and PAINT) super-resolution microscopy.
Utilizing short, interacting peptide tags for transient fluorescent protein binding.
Imaging of yeast membrane proteins that are intolerant to direct fluorescent protein fusion.
Employing orthogonal peptide interaction pairs for multiplexed imaging.

Main Results:

Successfully imaged yeast membrane proteins using LIVE-PAINT with minimal peptide tags (as short as 5 residues).
Demonstrated that LIVE-PAINT circumvents issues caused by direct fluorescent protein fusions.
Achieved concurrent nanoscale resolution of multiple distinct proteins using orthogonal peptide pairs.

Conclusions:

LIVE-PAINT is an effective super-resolution method for studying proteins that do not tolerate direct fluorescent tagging.
The technique enables high-resolution imaging of challenging cellular components, like yeast membrane proteins.
LIVE-PAINT offers a versatile platform for multiplexed nanoscale imaging in live cells.