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Updated: Jul 12, 2025
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Imaging Protein-protein Interactions in vivo
Published on: October 10, 2010
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Imaging Proteins Sensitive to Direct Fusions Using Transient Peptide-Peptide Interactions
Zoe Gidden1,2, Curran Oi3, Emily J Johnston1,4
1School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3DW, U.K.
Nano Letters
|November 2, 2023
Summary
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.
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.
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