Monitoring Proteome Stress in Live Cells Using HaloTag-Based Fluorogenic Sensor

Area of Science:

• Cellular Biology
• Biochemistry
• Molecular Biology

Background:

• Proteome stress, encompassing heat, osmotic, oxidative, and drug-induced conditions, significantly impacts cellular health.
• Monitoring cellular stress is crucial for understanding disease mechanisms and developing targeted therapeutics.
• Existing fluorescent sensors offer valuable insights but can be enhanced for broader applicability and robustness.

Purpose of the Study:

• To develop and validate a novel HaloTag-based fluorogenic sensor, termed AgHalo, for robust detection and quantification of proteome stresses.
• To demonstrate the sensor's utility in live bacterial and mammalian cells.
• To provide a versatile tool for dissecting cellular stress responses and identifying therapeutic interventions.

Main Methods:

• Development of a fluorogenic sensor utilizing the HaloTag protein system.
• Application of the AgHalo sensor in live bacterial and mammalian cell cultures.
• Quantitative analysis of proteome stress using fluorescence confocal microscopy and fluorescence plate readers.

Main Results:

• The AgHalo sensor successfully detected and quantified various proteome stresses in live cells.
• The sensor demonstrated robust performance across both bacterial and mammalian cell models.
• Quantitative fluorescence measurements provided reliable readouts of cellular stress levels.

Conclusions:

• The HaloTag-based AgHalo sensor is a powerful and versatile tool for monitoring proteome stresses in real-time within live cells.
• AgHalo facilitates the investigation of cellular stress mechanisms and the discovery of therapeutics for stress-related pathologies.
• This sensor system offers a valuable platform for both basic research and drug discovery efforts in cellular stress biology.