Sensitive detection and labelling of proteins with a molecular rotor-based fluorogenic probe

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Protein detection and labeling are crucial in biological research.
• Existing methods often require multiple steps or are not suitable for live-cell imaging.
• Development of sensitive and real-time probes is needed.

Purpose of the Study:

• To develop a sensitive fluorogenic reagent for protein detection and labeling.
• To demonstrate its utility in live-cell imaging and antibiotic susceptibility testing.

Main Methods:

• Design and synthesis of an environment-sensitive molecular rotor probe.
• Characterization of probe binding to protein amino residues versus free amino acids.
• Application of the probe for real-time imaging of membrane proteins in living cells.
• Utilizing the probe for determining the minimal inhibitory concentration (MIC) of antibiotics.

Main Results:

• The probe exhibits environment-sensitive fluorescence, significantly enhancing emission upon binding to proteins.
• The probe shows reversible, non-fluorescent interaction with free amino acids.
• Successful wash-free, real-time imaging of membrane proteins in living cells was achieved.
• Rapid determination of antibiotic MIC against bacteria was demonstrated.

Conclusions:

• The developed fluorogenic reagent is a sensitive and versatile tool for protein detection and labeling.
• Its ability for real-time, wash-free live-cell imaging opens new avenues in cell biology.
• The probe's utility in rapid MIC determination offers a valuable application in microbiology and drug discovery.