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A general chemical method to regulate protein stability in the mammalian central nervous system.

ABSTRACT: 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.

SUBMITTER: Iwamoto M

PROVIDER: S-EPMC2943492 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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A general chemical method to regulate protein stability in the mammalian central nervous system.

Iwamoto Mari M Björklund Tomas T Lundberg Cecilia C Kirik Deniz D Wandless Thomas J TJ

Chemistry & biology 20100901 9

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 protei ...[more]

PMID: 20851347

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