New, recombinant human Parkin purified from Sf21 cells!
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. 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. Parkin has been reported to generate poly-Ubiquitin chains in K6, K11, K48, and K63 linkages both in vitro and in vivo.
Recombinant human Parkin purified from insect cells by the method of Ordureau et al. is substantially more active than Parkin purified from bacteria. This recombinant protein is untagged.
- Ordureau, A. et al. (2015) PNAS 112: 6637-6642 (doi: 10.1073/pnas.1506593112)
- Ordureau, A. et al. (2014) Mol. Cell 56: 360-375 (doi: 10.1016/j.molcel.2014.09.007)