Active Parkinson Disease Protein 7 (PARK7)

Parkinson Disease Protein 7 (PARK7), a small multifunctional protein (20 kDa) containing 189 amino acids, which participates in transcriptional regulation  and mitochondrial regulation, and acts as a molecular chaperone, oxidative stress sensor, and glyoxalase. It has also been described as a protein and nucleotide deglycase. Mutations in Park7 are associated with a small percentage of hereditary early onset Parkinson’s disease. Leucine Rich Repeat Kinase 2 (LRRK2) has been identified as an interactor of PARK7, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human PARK7 and recombinant human LRRK2. Briefly, PARK7 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to LRRK2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PARK7 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human PARK7 and recombinant human LRRK2 was shown in Figure 1, the EC50 for this effect is 0.16 ug/mL.