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Updated: Sep 3, 2025
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Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
Published on: October 15, 2016
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Nanoscopic Spatial Association between Ras and Phosphatidylserine on the Cell Membrane Studied with Multicolor Super
Anna M Koester1, Kai Tao1, Malwina Szczepaniak1
1Program in Quantitative and Systems Biology, Department of Biomedical Engineering, Oregon Health & Science University, 2730 S Moody Ave., Portland, OR 97201, USA.
Biomolecules
|July 27, 2022
Summary
Ras GTPases interact with phosphatidylserine (PS) on cell membranes. New super-resolution imaging reveals isoform-specific PS-Ras associations, influenced by cellular context and cytoskeleton dynamics.
Area of Science:
- Cell Biology
- Molecular Biology
- Biophysics
Background:
- Ras small GTPases are key regulators of cellular signaling.
- Anionic lipid phosphatidylserine (PS) is crucial for membrane protein regulation.
- Previous studies suggest isoform-specific interactions between Ras and PS.
Purpose of the Study:
- To investigate the spatial relationship between Ras isoforms and PS in intact cells.
- To characterize the isoform-specific binding patterns of Ras GTPases to PS.
- To explore the influence of cellular context on PS-Ras interactions.
Main Methods:
- Utilized Lact-C2-GFP as a biosensor for PS.
- Employed multicolor super-resolution imaging based on DNA-PAINT.
- Performed spatial analysis of co-localization between Ras isoforms and PS.
Main Results:
- Revealed non-uniform distribution of PS on the cell membrane, co-enriching with caveolae.
- Demonstrated that both KRas-G12D and HRas-G12V co-enrich with PS in U2OS cells.
- Observed distinct association patterns: HRas-G12V consistently co-enriched with PS, while KRas-G12D showed variable co-enrichment.
- Showed differential impact of actin cytoskeleton perturbations on PS-Ras isoform associations.
Conclusions:
- Ras-phosphatidylserine association is context-dependent and isoform-specific.
- Multiplexed super-resolution imaging is a powerful tool for studying Ras-membrane interactions.
- Findings provide insights into the complex interplay between Ras GTPases and the cell membrane.
