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A General Chemical Method to Regulate Protein Stability in the Mammalian Central Nervous System

2010; Elsevier BV; Volume: 17; Issue: 9 Linguagem: Inglês

10.1016/j.chembiol.2010.07.009

1879-1301

Mari Iwamoto, Tomas Björklund, Cecilia Lundberg, Deniz Kirik, Thomas J. Wandless,

Endoplasmic Reticulum Stress and Disease

The ability to make specific perturbations to biological molecules in a cell or organism is a central experimental strategy in modern research biology. We have developed a general technique in which the stability of a specific protein is regulated by a cell-permeable small molecule. Mutants of the Escherichia coli dihydrofolate reductase (ecDHFR) were engineered to be degraded, and, when this destabilizing domain is fused to a protein of interest, its instability is conferred to the fused protein resulting in rapid degradation of the entire fusion protein. A small-molecule ligand trimethoprim (TMP) stabilizes the destabilizing domain in a rapid, reversible, and dose-dependent manner, and protein levels in the absence of TMP are barely detectable. The ability of TMP to cross the blood-brain barrier enables the tunable regulation of proteins expressed in the mammalian central nervous system.