A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules.

Area of Science:

• Molecular Biology
• Biochemistry
• Cell Biology

Background:

• * Precise control over protein function is crucial for understanding complex biological systems.
• * Existing methods for protein perturbation often lack speed, reversibility, or specificity.

Purpose of the Study:

• * To develop a general and controllable method for regulating protein stability in mammalian cells.
• * To enable conditional perturbation of specific proteins using synthetic molecules.

Main Methods:

• * Engineering destabilizing mutants of the human FKBP12 protein.
• * Fusing these destabilizing domains to target proteins of interest.
• * Utilizing cell-permeable synthetic ligands to control protein degradation.

Main Results:

• * Engineered FKBP12 mutants confer rapid and constitutive degradation to fused proteins.
• * Addition of a synthetic ligand reverses degradation, restoring protein function.
• * The system demonstrates specificity, speed, reversibility, and dose-dependence.

Conclusions:

• * This novel strategy provides unprecedented control over protein stability.
• * The method facilitates conditional protein perturbation in various experimental settings.
• * It offers a powerful tool for dissecting biological pathways.