Recombinant DJ - 1 (PARK7) Protein, Human, GST tagged (GST - DJ - 1)
Greater than 95% as determined by SDS-PAGE.
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. GST tag was not removed. The molecular weight of the recombinant GST-DJ-1protein is 47.3 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 mutated1-5. Recently it was demonstrated that DJ-1 can act as an atypical peroxiredoxin-like peroxidase5 and poses protease activity4.
Figure 1. Human GST-DJ-1 on SDS-PAGE. Purified GST-DJ-1 was loaded onto 10-20% Tris-H gel at 3 μg/well and resolved at 200V for 40 min. 1=Uninduced cell lysate, 2=Induced cell lysate,3=Purified GST-DJ-1 protein.
Figure 2. Western Immunoblot of DJ-1. Purified DJ-1 protein was probed with anti-DJ-1 specific antibody.
Figure 3.GST-DJ-1 Proteolytic Activity. GST-DJ-1 was loaded at 200 μg/well with 1mM DTT and assayed for proteolytic activity using Green Protease Assay Kit (AnaSpec Cat. # AS-71124).
The purified GST-DJ-1 is supplied as sterile and frozen at 1 mg/ml in 50 mM Tris-Cl/150mM NaCl, pH=8.0. Store at 2-4 ºC for immediate use within 1-2 weeks or at –80 ºC for up to 12 months. Avoid repeated freeze-thaw cycles.
1. Huai Q., et. al., FEBS Letters v549 (2003): 171-175 2. Inden M., et. al., J. of Pharma. Sci. v117 (2011): 189-203 3. Kitamura Y., et. al., Mol. Neurodeg. v6 n48 (2011): 1-19 4. Chen J., et. al., Hum. Mol. Genetics v19 n12 (2010): 2395-2408 5. Andres-Mateos E., et. al., Proc. Nat. Acad. Sci. v104 (2007): 14807-14812