Synthesis and analysis of stabilizing ligands for FKBP-derived destabilizing domains.
We recently identified mutants of the human FKBP12 protein that are unstable and rapidly degraded when expressed in mammalian cells. We call these FKBP mutants destabilizing domains (DDs), because their instability is conferred to any protein fused to the DDs. A cell-permeable ligand binds tightly to the DDs and prevents their degradation, thus providing small molecule control over intracellular protein levels. We now report the synthesis and functional characterization of a stabilizing ligand called Shield-2. The synthesis of Shield-2 is efficient, and this ligand binds to the FKBP(F36V) protein with a dissociation constant of 29 nM.
Grimley
Joshua S
JS
Department of Chemical & Systems Biology, Stanford University, 318 Campus Drive, Clark Center W350A, Stanford, CA 94305-5441, USA.
Chen
Denise A
DA
Banaszynski
Laura A
LA
Wandless
Thomas J
TJ
D000818:Animals; D006801:Humans; D008024:Ligands; D051379:Mice; D022021:Tacrolimus Binding Proteins / Q000737:chemistry*
D008024:Ligands; D022021:Tacrolimus Binding Proteins