Ubiquitin carboxyl-terminal hydrolase 30 (USP30) is a member of the C19 family of peptidases and is tethered to the outer mitochondrial surface via a trans-membrane domain. USP30 activity is antagonistic to the pro-mitophagy signal generated by the activities of E3 Ubiquitin Ligase Parkin (PARK2) and kinase PINK1, which are stimulated by mitochondrial damage. In vivo, USP30 cleaves Ubiquitin from mitochondrial surface proteins such as MIRO1, TOMM20, MFN1, and MFN2. The recombinant enzyme displays a strong preference for K6- and K11-linked polyubiquitin chains in vitro, but has substantially lower activity on polyubiquitin chains that have been phosphorylated with PINK1 kinase. This recombinant protein contains amino acids 57-517 of the full-length protein, and a C-terminal 6-His tag.
Use & Storage
Reaction conditions will need to be optimized for each specific application. We recommend an initial USP30 concentration of 20-100 nM in reactions utilizing Ubiquitin-AMC or Ubiquitin-Rhodamine substrates (U-550 or U-555), or 200-500 nM when digesting recombinant poly-Ubiquitin chain substrates.
Store at -80°C. Avoid multiple freeze/thaw cycles.