Parkin mitochondrial recruitment assay from Innoprot is based of localization changes of two fluorescent proteins; the Parkin fluorescent fusion polypeptide that changes its localization from the cytosol to the outer mitochondrial membrane (OMM); and the
other fluorescent fusion polypeptide (MTS-tGFP), which binds directly to OMM, allowing the mitochondrial integrity analysis.
Several compounds promote Parkin recruitment, but also generate stress and increase cell toxicity. So a good, promising positive compound would be one that promotes Parkin binding to mitochondria to accelerate Parkin protective function but without deeply altering mitochondrial integrity. Our model allows the identification of compounds that can promote the mitochondrial localization of Parkin without severe mitochondrial damage induction.
Cell line used: PARK2-FP602+MTS-TGFP / U2OS cell line
Readout: Parkin recruitment by mitochondria
Agonist: Proscillaridin A
EC50 Agonist: 2.59 × 10-8 M
Type of assay: Cell based / Functional
Detection Method: Image Analysis