Visible-Light-Controlled Lysine-Selective Crosslinking Decodes Protein Complexes and Dynamic Interactomes in Live...

Area of Science:

• Chemical biology
• Proteomics
• Molecular and systems biology

Background:

• Protein complexes and protein-protein interactions (PPIs) are crucial for cellular functions.
• Existing crosslinking strategies face challenges in analyzing dynamic PPIs in native biological settings.

Purpose of the Study:

• To develop a novel photo-crosslinking strategy for precise analysis of protein complexes and dynamic interactomes in live cells.
• To introduce Visible-light-controlled Lysine-selective crosslinking (VL-XL) for temporal control and high biocompatibility.

Main Methods:

• Development of visible-light-inducible, lysine-specific homobifunctional photo-crosslinkers.
• Application of the VL-XL strategy for residue-specific crosslinked peptide identification.
• Integration with mass spectrometry (MS) for data analysis.

Main Results:

• VL-XL enables high-confidence data acquisition and streamlined MS analysis.
• Successfully profiled the time-resolved, epidermal growth factor (EGF)-stimulated epidermal growth factor receptor (EGFR) interactome.
• Discovered neo-substrates, such as Sestrin-2 (SESN2), in molecular glue degrader-induced E3 ligase interactomes.

Conclusions:

• VL-XL is a robust chemical tool for in-depth analysis of protein complexes and dynamic interactomes.
• Provides valuable insights into EGFR signaling regulation and E3 ligase interactions.
• Opens innovative avenues for identifying novel therapeutic targets for protein degradation.