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Updated: Jan 24, 2026
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Characterization of Neuronal Lysosome Interactome with Proximity Labeling Proteomics
Published on: June 23, 2022
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The EMARS Reaction for Proximity Labeling.
Koichi Honke1,2, Arisa Miyagawa-Yamaguchi3, Norihiro Kotani4
1Department of Biochemistry, Kochi University Medical School, Nankoku, Kochi, Japan. khonke@kochi-u.ac.jp.
Methods in Molecular Biology (Clifton, N.J.)
|May 25, 2019
Summary
We developed a new proximity labeling method using enzyme-mediated activation of radical source (EMARS) to identify proteins interacting with a molecule of interest in living cells.
Area of Science:
- Cellular Biology
- Molecular Biology
- Biochemistry
Background:
- Understanding cellular processes requires mapping protein-protein interactions within living cells.
- Existing methods may have limitations in spatial resolution or applicability.
Purpose of the Study:
- To develop and describe a novel proximity labeling technique for identifying proteins near a molecule of interest in live cells.
- To provide protocols for reagent preparation, in-cell labeling, and protein identification.
Main Methods:
- Utilized an enzyme-mediated activation of radical source (EMARS) reaction.
- Employed horseradish peroxidase (HRP) to generate radicals from labeling reagents.
- Covalently tagged proximal proteins to a molecule of interest.
Main Results:
- Successfully developed a proximity labeling method applicable in living cells.
- Established protocols for preparing labeling reagents and performing the labeling procedure.
- Enabled identification of neighboring proteins through covalent tagging.
Conclusions:
- The EMARS-based proximity labeling method is effective for mapping protein interactions in situ.
- This technique facilitates molecular-level understanding of cellular processes.
- The described protocols offer a valuable tool for cell biology research.
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