Small-molecule displacement of a cryptic degron causes conditional protein degradation

KM Bonger, L Chen, CW Liu, TJ Wandless - Nature chemical biology, 2011 - nature.com
KM Bonger, L Chen, CW Liu, TJ Wandless
Nature chemical biology, 2011nature.com
The ability to rapidly regulate the functions of specific proteins in living cells is a valuable
tool for biological research. Here we describe a new technique by which the degradation of
a specific protein is induced by a small molecule. A protein of interest is fused to a ligand-
induced degradation (LID) domain, resulting in the expression of a stable and functional
fusion protein. The LID domain is comprised of the FK506-and rapamycin-binding protein
(FKBP) and a 19-amino-acid degron fused to the C terminus of FKBP. In the absence of the …
Abstract
The ability to rapidly regulate the functions of specific proteins in living cells is a valuable tool for biological research. Here we describe a new technique by which the degradation of a specific protein is induced by a small molecule. A protein of interest is fused to a ligand-induced degradation (LID) domain, resulting in the expression of a stable and functional fusion protein. The LID domain is comprised of the FK506- and rapamycin-binding protein (FKBP) and a 19-amino-acid degron fused to the C terminus of FKBP. In the absence of the small molecule Shield-1, the degron is bound to the FKBP fusion protein and the protein is stable. When present, Shield-1 binds tightly to FKBP, displacing the degron and inducing rapid and processive degradation of the LID domain and any fused partner protein. Structure-function studies of the 19-residue peptide showed that a 4-amino-acid sequence within the peptide is responsible for degradation.
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