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Updated: Feb 15, 2026
08:59
4D Imaging of Protein Aggregation in Live Cells
Published on: April 5, 2013
17.8K
A Fluorescence-Based Sensor Assay that Monitors General Protein Aggregation in Human Cells.
Marisa Pereira1, Diogo Tomé1, Ana S Domingues1
1iBiMED - Institute of Biomedicine Department of Medical Sciences University of Aveiro, 3810-193 Aveiro, Portugal.
Biotechnology Journal
|January 19, 2018
Summary
Researchers developed a new method using Heat Shock Protein 27 (HSP27) fused to Green Fluorescent Protein (GFP) to monitor protein aggregation in human cells. This tool aids in identifying genes involved in protein misfolding diseases.
Area of Science:
- Cell Biology
- Biochemistry
- Molecular Biology
Background:
- Protein conformational disorders arise from misfolding and aggregate accumulation.
- Monitoring protein aggregation in living cells is crucial for understanding these diseases.
Purpose of the Study:
- To develop a sensitive and simple method for monitoring general protein aggregation in human cells.
- To validate a reporter system for detecting protein misfolding and aggregation.
Main Methods:
- Constructed a HeLa stable cell line expressing a Heat Shock Protein 27:Green Fluorescent Protein (HSP27:GFP) chimeric reporter.
- Validated the reporter system by exposing cells to proteostasis-interfering agents (Arsenite, MG132, Aβ-peptide).
Main Results:
- The HSP27:GFP reporter successfully detected and monitored protein aggregation in living cells.
- Exposure to proteome destabilizers induced re-localization of HSP27:GFP fluorescence to foci.
Conclusions:
- The HSP27:GFP reporter system is a functional tool for detecting and following protein aggregation in real-time.
- This reporter system can facilitate wide-genetic screens to identify genes and pathways involved in protein misfolding and aggregation.
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