Inducible secretion of proteins

Inducible secretion of proteins. Fusion proteins containing DmrD domains localize to the endoplasmic reticulum as massive aggregates (left). When the D/D Solubilizer is added, it dissolves the aggregates and allows the protein to be exported through the secretory apparatus (right). To ensure secretion of the authentic protein, a furin cleavage site is positioned between the DmrD domains and the protein of interest. Since furin is exclusively expressed in the trans Golgi, the fusion protein will be processed as it traverses this compartment, resulting in the secretion of the correctly processed protein. This method has been used to induce rapid, transient and tightly regulated secretion of human growth hormone (hGH) and insulin [Rivera, V. M., et al. (2000) Science 287(5454):826–830].

Molecular structure of the D/D Solubilizer

Molecular structure of the D/D Solubilizer. The D/D Solubilizer performs the same function as the AP21998 ligand. It is a different molecule than AP21998.

Secretion of DmrD-tagged luciferase after addition of D/D Solubilizer

Secretion of DmrD-tagged luciferase after addition of D/D Solubilizer. 7 hr after transfection with a DmrD-tagged Metridia luciferase construct, cells were split into wells of a 6-well plate. The medium was removed and fresh medium was added containing increasing concentrations of D/D Solubilizer. 18 hr later, the media was collected and analyzed using our Ready-To-Glow™ Secreted Luciferase Reporter System (Cat. No. 631731).

Molecular weights of iDimerize ligands

Molecular weights of iDimerize ligands.

Rivera, V. M. et al. Regulation of protein secretion through controlled aggregation in the endoplasmic reticulum. Science 287, 826–30 (2000).

Volchuk, A. et al. Megavesicles Implicated in the Rapid Transport of Intracisternal Aggregates across the Golgi Stack. Cell 102, 335–348 (2000).