Recombinant Human Parkin Protein, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Parkin Protein, CF Summary

• Details of Functionality:

  Reaction conditions will need to be optimized for each specific application. As supplied, Parkin has negligible E3 ligase activity as determined by the lack of autoubiquitination in an in vitro assay. Parkin ligase activity is greatly enhanced by phosphorylation with PINK1 kinase (AP-180) or by the addition of low concentrations of phosphorylated Ubiquitin (U-102).

• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Parkin protein
• Accession #: O60260
• Protein/Peptide Type: Recombinant Enzymes
• Gene: PARK2
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Applications/Dilutions

• Theoretical MW: 52 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
• Buffer: X mg/ml (X μM) in 25 mM Tris pH 8.5, 200 mM NaCl, 0.03% Brij35, 10% (v/v) Glycerol, 5 mM TCEP
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Parkin Protein, CF

• AR-JP
• E3 ubiquitin ligase
• E3 ubiquitin-protein ligase parkin
• EC 6.3.2.-
• LPRS2
• PARK2
• parkin 2
• Parkin
• Parkinson disease protein 2
• Parkinson juvenile disease protein 2
• parkinson protein 2, E3 ubiquitin protein ligase (parkin)
• PDJ
• PDJjuvenile) 2, parkin
• PRKN

Background

The E3 Ubiquitin ligase Parkin (encoded by the PARK2 gene) is an essential part of the cellular machinery that participates in the removal of damaged mitochondria. Mutations in PARK2 are known to cause a form of Parkinson's disease known as autosomal recessive juvenile Parkinson's disease (AR-JP), and the mechanisms by which defective Parkin ligase contributes to the dopaminergic cell death in this disease is an area of intense investigation.

Reported substrates for Parkin include BCL2, GPR37, MIRO1, MFN1, MFN2, TOMM20, USP30, and many others. Parkin (an RBR-class Ubiquitin ligase) structures have recently been reported by multiple groups, and reveal that the ligase is folded upon itself to produce an auto-inhibited state. The auto-inhibition is relieved by interactions with PINK1 kinase (which can phosphorylate both Parkin and Ubiquitin at serine residue number 65) and pS65 phospho-Ubiquitin by mechanisms that are under investigation.

In vitro, Parkin may be activated by treatment with recombinant PINK1, or addition of low concentrations of pS65-phosphoubiquitin. Parkin has been reported to generate poly-Ubiquitin chains in K6, K11, K48, and K63 linkages both in vitro and in vivo. This recombinant protein is untagged.

1. Bingol, B. et al. (2014) Nature 510: 370
2. Ordureau, A. et al. (2014) Mol. Cell 56: 360
3. Riley, B.E. et al. (2013) Nat. Comm. 4: doi:10.1038/ncomms2982
4. Saraff, S.A. et al. (2013) Nature 496: 372
5. Spratt, D.E. et al. (2013) Nat. Comm. 4: doi:10.1038/ncomms2983
6. Trempe, J.F. et al. (2013) Science 340: 1451
7. Wauer T. et. al. (2015) Nature 524: 370
8. Wauer T. & Komander, D. (2013) EMBO J 32: 2099

Bioinformatics

• Gene Symbol: PARK2
• Uniprot:
  • O60260(Human)