DJ-1 (PARK7) Protein

The sequence (Accession # NP_001116849) corresponding to the full length human DJ-1 protein along with N-terminal GST tag was expressed in E. coli. The recombinant DJ-1 protein was purified from bacterial lysate using GST affinity chromatography followed by GST tag cleavage and removal. The molecular weight of the recombinant DJ-1protein is 19.9 kDa. DJ-1 (also known as PARK7, Parkinson disease protein 7) is a 189 amino acid protein that is ubiquitously expressed in many organs including brain, liver, kidney, pancreas, heart, and others1-5. Although DJ-1 was originally discovered as a novel oncogene product, it was found to play several other roles in biological processes. They include regulation of RNA binding activity, fertility, anti-oxidative stress, and linked to the early onset of Parkinson disease when mutated 1-5. Recently it was demonstrated that DJ-1 can act as an atypical peroxiredoxin-like peroxidase5 and possesses protease activity4. Figure 1. Human DJ-1 on SDS-PAGE. Purified DJ-1 was loaded onto 10-20% Tris-HCl gel at 3 ug/well and resolved at 200V for 40 min. 1=Uninduced cell lysate, 2=Induced cell lysate, 3=Purified and cleaved DJ-1 protein. Figure 2. Western Immunoblot of DJ-1. Purified DJ-1 protein was probed with anti-DJ-1 specific antibody. Figure 3. DJ-1 Proteolytic Activity. DJ-1 was loaded at 200 ug/well with 1mM DTT and assayed for proteolytic activity using Green Protease Assay Kit (AnaSpec Cat. # AS-71124).

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