Protein-Protein Interaction in Multicomponent Reaction Enables Chemoselective, Site-Selective, and Modular Labeling...

Area of Science:

• Chemical Biology
• Protein Engineering
• Biochemistry

Background:

• Single-site protein modification is challenging due to diverse protein functionalities.
• Existing methods often lack specificity or are difficult to implement.
• Controlling modification sites is crucial for understanding protein function and engineering new ones.

Purpose of the Study:

• To develop a novel method for achieving site-selective protein modification.
• To leverage protein-protein interactions (PPI) for directing chemical modifications.
• To enable late-stage installation of mono- and dual-probes onto proteins.

Main Methods:

• Utilized chemoselective, reversible generation of reactive intermediates.
• Employed protein-protein interaction specificity to guide irreversible modification.
• Applied a multicomponent aza-Morita-Baylis-Hillman (aza-MBH) reaction driven by dis(in)tegrate (DIN) theory.

Main Results:

• Achieved homogeneous and modular single-site protein modification.
• Demonstrated the ability to install mono- and dual-probes at a specific site.
• Successfully harnessed PPI to drive site-selective chemical reactions.

Conclusions:

• The developed method offers a powerful strategy for precise protein engineering.
• This approach overcomes limitations of traditional single-site modification techniques.
• The DIN theory-driven aza-MBH reaction provides a versatile platform for protein functionalization.