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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

Predicted Molecular Mass

52 kDa

Product Datasheets

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

E3-160

Formulation	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
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Reconstitution Calculator

Background: Parkin

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.

References

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

Long Name

Parkinson Disease [autosomal recessive, juvenile] 2, Parkin [PARK2], transcript variant 1

Entrez Gene IDs

5071 (Human); 50873 (Mouse); 56816 (Rat)

Alternate Names

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