• Purity

  >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie? Blue stain.

• Activity

  Recombinant Human USP46/UAF1 complex is a Ubiquitin-specific deconjugating enzyme heterodimer.  Reaction conditions will need to be optimized for each specific application.  We recommend an initial USP46/UAF1 Complex concentration of 20-200 nM when using Ubiquitin-AMC or Ubiquitin-Rhodamine (U-550, U-555) as a substrate.  In vitro, USP46/UAF1 complex does not efficiently hydrolyze poly-Ubiquitin chains in the absence of WDR20.

• Source

  Spodoptera frugiperda,     Sf 21 (baculovirus)-derived Contains a C-terminal 6-His tag

• Accession #

• Predicted Molecular Mass

  43 kDa (USP46), 77 kDa (UAF1)

Background: USP46/UAF1 Complex

Ubiquitin carboxyl-terminal hydrolase 46 (USP46) is a specialized cysteine protease with a predicted molecular weight of 42 kDa. USP46 is a member of the peptidase C19 family and USP12/USP46 subfamily, and the human protein shares 100% amino acid sequence identity with its mouse ortholog. Like USP1 and USP12, USP46 lacks appreciable activity in the absence of its co-activator UAF1. Addition of UAF1 increases the in vitro activity of USP46 10-fold or more.  Biologically, USP46 has been implicated in the regulation of Alpha-Amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid receptors (AMPARs). USP46 is able to deubiquitinate AMPARs both in vivo and in vitro, and by regulating the status of AMPAR ubiquitination and turnover, this deubiquitinase may play an important role in synaptic plasticity and brain function. Recombinant USP46 contains a C-terminal 6-His tag.  Recombinant UAF1 also contains a C-terminal 6-His tag.  The assembled complex contains 25 μg USP46 and 45 μg UAF1, resulting in a 1:1 stoichiometry.

• References:

1. Cohn, M.A. et al. (2009) J Biol Chem. 284: 5343

2. Huo, Y. et al. (2015) J Neurochem. doi: 10.1111/jnc.13194.

3. Ohashi, M. et al. (2015) PLoS Pathog.  DOI: 10.1371/journal.ppat.1004822

• Long Name:

  Ubiquitin Specific Protease 46/USP1-associated Factor 1 Complex

• Alternate Names:

  USP46/UAF1 Complex