A versatile synthetic dimerizer for the regulation of protein–protein interactions

Abstract

The use of low molecular weight organic compounds to induce dimerization or oligomerization of engineered proteins has wide-ranging utility in biological research as well as in gene and cell therapies. Chemically induced dimerization can be used to activate intracellular signal transduction pathways or to control the activity of a bipartite transcription factor. Dimerizer systems based on the natural products cyclosporin, FK506, rapamycin, and coumermycin have been described. However, owing to the complexity of these compounds, adjusting their binding or pharmacological properties by chemical modification is difficult. We have investigated several families of readily prepared, totally synthetic, cell-permeable dimerizers composed of ligands for human FKBP12. These molecules have significantly reduced complexity and greater adaptability than natural product dimers. We report here the efficacies of several of these new synthetic compounds in regulating two types of protein dimerization events inside engineered cells--induction of apoptosis through dimerization of engineered Fas proteins and regulation of transcription through dimerization of transcription factor fusion proteins. One dimerizer in particular, AP1510, proved to be exceptionally potent and versatile in all experimental contexts tested.

Synthetic Multivalent Ligands as Probes of Signal Transduction

“…[192] Such strategies have been expanded to include heterodimerizers and dimerizers that avoid endogenous proteins (e.g., FKBP) and specifically target engineered fusion proteins. [192][193][194] As discussed above, both integrin proximity and conformation contribute to the switch between low-and high-affinity integrin binding. To dissect the role of conformational change from that of receptor proximity, a CID strategy was employed.…”

Protein‐protein interactions as a target for drugs in proteomics

“…Now the most prevalent dimerizer is FK1012 (nontoxic lipid-soluble dimeric form of the drug FK506), but other anchor groups are also used [108,109,[118][119][120]. Linkers consist of 5-16 atoms [121,122]. The concentrations required for induction of dimerization by such compounds vary in a relatively narrow range of 1-10 nM [121,123].…”

“…Linkers consist of 5-16 atoms [121,122]. The concentrations required for induction of dimerization by such compounds vary in a relatively narrow range of 1-10 nM [121,123]. The effectiveness of dimerizers depends on the anchors affinity [122] and the length of the linkers [121,122].…”

“…At present these systems are used only in the laboratory. Dimerizers can induce cell proliferation [124], transcription [118,121] and apoptosis [121,123]. It is proposed to use these systems in gene therapy.…”

“…It is proposed to use these systems in gene therapy. Potentially, in clinical practice dimerizers can be used for induction of cell proliferation [124], or for elimination of the transferred gene product and genetically modified cells by dimerizer-inducing apoptosis [121,123].…”