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Published on: June 24, 2017
Kinetic and Spatial Resolution by dSTORM of miniSOG-Mediated Proximity Labeling in Mammalian Cells
Benoît Arnould1, Alexandria L Quillin1, Aastha1
1Department of Chemistry, Washington University in St. Louis, St. Louis, Missouri, USA.
Chembiochem : a European Journal of Chemical Biology
|July 8, 2026
Summary
The mini Singlet Oxygen Generator (miniSOG) allows proximity labeling with blue light. This study quantifies its cellular labeling range and kinetics, revealing tunable parameters for proteomic and transcriptomic applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- The mini Singlet Oxygen Generator (miniSOG) is a blue-light activated protein for photocatalytic proximity labeling.
- Its application in proteomics and transcriptomics is widespread, but kinetic and spatial parameters in cells are not well-defined.
Purpose of the Study:
- To quantitatively characterize the in situ kinetic and spatial parameters of miniSOG activity in mammalian cells.
- To establish a framework for optimizing miniSOG-based experimental design.
Main Methods:
- Utilized a microscope-based irradiation platform combined with single-molecule localization microscopy (SMLM).
- Analyzed miniSOG activity in fixed mammalian cells under varying blue-light illumination conditions.
- Quantified labeling kinetics and effective spatial reach using super-resolution imaging.
Main Results:
- Identified an optimal intermediate blue-light power for sustained miniSOG activity and signal accumulation.
- Observed photoinactivation at higher illumination power levels.
- Demonstrated that the effective labeling diameter expands with illumination time (approx. 70 nm to 180 nm over 30 min).
- Showed miniSOG spatial reach is consistent across cellular compartments and sensitive to singlet oxygen quenching.
Conclusions:
- Provided the first in situ quantification of miniSOG's kinetic and spatial capture parameters.
- Established that miniSOG's effective proximity labeling range is tunable for diverse biological applications.
- Emphasized the importance of understanding these parameters for accurate interpretation of miniSOG-based proteomic and transcriptomic studies.
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