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Updated: Jan 31, 2026
07:38
Fluorescent Labeling of COS-7 Expressing SNAP-tag Fusion Proteins for Live Cell Imaging
Published on: May 17, 2010
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A Fluorogenic AggTag Method Based on Halo- and SNAP-Tags to Simultaneously Detect Aggregation of Two Proteins in Live
Kwan Ho Jung1, Sojung F Kim1, Yu Liu1
1Department of Chemistry, The Pennsylvania State University, University Park, PA, 16802, USA.
Chembiochem : a European Journal of Chemical Biology
|January 5, 2019
Summary
Researchers developed a new method to visualize two different protein aggregates simultaneously in living cells. This advancement in protein aggregation research aids in understanding neurodegenerative diseases.
Area of Science:
- Cell Biology
- Biochemistry
- Neuroscience
Background:
- Protein aggregation is implicated in neurodegenerative diseases.
- Current tools lack the ability to visualize multiple protein aggregation events in live cells simultaneously.
- Previous Aggregation Tag (AggTag) method using Halo-tag detected only one protein of interest (POI) aggregation at a time.
Purpose of the Study:
- To expand the AggTag method for simultaneous visualization of two different POIs.
- To develop a new AggTag probe for biorthogonal detection of protein aggregation.
- To enable simultaneous tracking of distinct pathogenic protein aggregates within the same cell.
Main Methods:
- Developed a new AggTag probe (P2) utilizing SNAP-tag technology and a green solvatochromic fluorophore.
- Fused POIs with SNAP-tag and Halo-tag ligands.
- Employed confocal imaging and chemical crosslinking experiments.
- Utilized orthogonal fluorescence of P1 (Halo-tag based) and P2 (SNAP-tag based) probes.
Main Results:
- The new P2 probe successfully reported on both soluble oligomers and insoluble aggregates of SNAP-tagged POIs in live cells.
- Demonstrated the ability to visualize aggregation of two distinct POIs concurrently within the same cell.
- Confirmed the fluorogenic and biorthogonal nature of the expanded AggTag system.
Conclusions:
- The expanded AggTag method using SNAP-tag technology enables simultaneous, fluorogenic, and biorthogonal detection of two distinct protein aggregation events in live cells.
- This provides a powerful new tool for studying the complex mechanisms of protein aggregation in diseases.
- Facilitates a deeper understanding of cellular processes involving multiple aggregating proteins.
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