Covalent Solvatochromic Proteome Stress Sensor Based on the Schiff Base Reaction

Area of Science:

• Chemical Biology
• Biochemistry
• Proteomics

Background:

• Covalent probes for detecting aggregated proteins are scarce.
• Existing methods struggle to selectively target and analyze protein aggregates.
• Understanding protein aggregation is crucial for disease research.

Purpose of the Study:

• To develop and characterize novel covalent solvatochromic probes for aggregated proteins.
• To investigate the Schiff base reaction for selective protein modification.
• To visualize and quantify features of aggregated proteomes.

Main Methods:

• Design and synthesis of covalent solvatochromic probes (e.g., P1 probe with aldehyde).
• Utilizing Schiff base chemistry for selective covalent modification of lysine residues.
• Biochemical assays, mass spectrometry, and fluorescence spectroscopy for analysis.
• Exploiting solvatochromism for imaging and quantification.

Main Results:

• The P1 probe selectively modifies aggregated proteins over folded ones via Schiff base reaction.
• Protein aggregation leads to enhanced fluorescence and a significant blue shift.
• The probes demonstrate generality across different protein scaffolds.
• Visualized distinct morphologies and quantified polarity heterogeneity within aggregates.

Conclusions:

• Novel covalent probes enable selective detection and modification of aggregated proteins.
• Schiff base chemistry is a viable strategy for targeting protein aggregates.
• The developed probes facilitate visualization and characterization of protein aggregation.
• This approach opens new avenues for studying difficult-to-analyze protein aggregates.