Genetically encoded bioorthogonal tryptophan decaging in living cells

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Tryptophan (Trp) is crucial for protein structure and function due to its unique chemical properties.
• Controlling Trp interactions in vivo is challenging but essential for protein manipulation.
• Existing methods lack generalizability and site-specific control for Trp interactions.

Purpose of the Study:

• To develop a generalizable platform for blocking and rescuing Trp interactions in proteins.
• To enable gain-of-function manipulation of Trp-containing proteins in vivo.
• To achieve site-specific activation of Trp on proteins of interest in living cells.

Main Methods:

• Genetically encoding an N1-vinyl-caged Trp (Trp-CAGE).
• Utilizing an optimized inverse electron-demand Diels-Alder reaction for rapid and bioorthogonal decaging.
• Applying the Trp-CAGE strategy to various protein families, including catalase-peroxidases and kinases.

Main Results:

• Demonstrated rapid and bioorthogonal decaging of Trp-CAGE in living cells.
• Successfully activated Trp site-specifically in diverse protein families.
• Showcased temporal control over epigenetic signaling modulation.
• Identified over 28,000 candidate proteins for bioorthogonal Trp activation.

Conclusions:

• The Trp-CAGE strategy provides a powerful tool for precise, bioorthogonal Trp activation.
• This method enables the modulation of protein interactions and functions in native cellular environments.
• The platform has broad applicability for manipulating diverse proteins and understanding cellular processes.