Takara iDimerize Reverse Dimerization System (inducible secretion)

Takara iDimerize Reverse Dimerization System (Inducible Secretion)

The iDimerize Reverse Dimerization System enables real-time, small-molecule control over the disruption of protein complexes. A protein of interest is fused to the DmrD binding domain, and the fusion protein molecules aggregate unless the D/D Solubilizer ligand is present. Both plasmid and lentiviral (Lenti-X) vector formats are available.

Reverse Dimerization: Disrupting Protein-Protein Interactions

The iDimerize Reverse Dimerization System is a “reverse dimerization” system—aggregation is the resting state, and the D/D Solubilizer breaks up protein-protein interactions. This system complements inducible dimerization and can be used analogously to create inducible alleles. Processes that are activated by oligomerization can be turned off using this kit, allowing reversible control of protein activity.

Inducible Secretion

The ability to generate large protein aggregates provides unique applications. For instance, fusion proteins with a secretory signal sequence can be stored as aggregates in the endoplasmic reticulum. Upon addition of the ligand, a rapid pulse of protein secretion can be induced. This method has been used to achieve tightly regulated secretion of human growth hormone (hGH) and insulin (Rivera et al., 2000).

Protein aggregates are also valuable in protein trafficking studies. This approach has led to discoveries such as “mega-vesicles” transporting cargo across the Golgi stack (Volchuk et al., 2000).

D/D Solubilizer Ligand

The D/D Solubilizer is a synthetic, cell-permeable ligand used to disrupt dimerization of fusion proteins containing the DmrD domain. It has been tested in vitro and in mice and found to be nontoxic. Optimal conditions can be determined by testing various concentrations (10–500 nM) and incubation times (30 minutes to 12+ hours) for a complete dose-response profile.

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