Enabling Protein Degradation Drug Discovery

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  • Name
    Catalogue Number
    Size
    Price
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  • Name:
    UBE2R2 (CDC34B) [6His-tagged]
    Catalogue Number:
    62-0094-020
    Size:
    20 µg
    Price:
    £100
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  • Species
    Human
  • Source
    E. coli expression
  • Quantity
    20 μg
  • Storage
    -70°C
  • Concentration
    1 mg/ml
  • Formulation
    50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol
  • Molecular Weight
    ~31 kDa
  • Stability
    12 months at -70°C; aliquot as required
  • Protein Sequence
    Accession number: AAH04862. For full protein sequence information download the Certificate of Analysis pdf.
  • QA; Protein Identification
    Confirmed by mass spectrometry.
  • QA; Activity
    E2-Ubiquitin Thioester Loading Assay: The activity of His-UBE2R2 was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the His-UBE2R2 E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and His-UBE2R2 enzymes in the presence of ubiquitin and ATP at 30°C was compared at two time points, T0 and T10 minutes. Sensitivity of the ubiquitin/His-UBE2R2 thioester bond to the reducing agent DTT was confirmed.

The enzymes of the ubiquitylation pathway play a pivotal role in a number of cellular processes including the regulated and targeted proteasomal degradation of substrate proteins. Three classes of enzymes are involved in the process of ubiquitylation; activating enzymes (E1s), conjugating enzymes (E2s) and protein ligases (E3s). UBE2R2 is a member of the E2 conjugating enzyme family and cloning of the human gene was first described by Semplici et al. (2002). Site directed mutagenesis studies have shown that serine 233 in the C-terminal domain of UBE2R2 is the site at which CK2-dependent phosphorylation occurs (Semplici et al., 2002). In vitro binding experiments have also demonstrated that phosphorylated UBE2R2 and UBE2R1 bind specifically to the F-box protein beta-TRCP, which results in enhanced degradation of beta-catenin (a substrate of the Beta Transducin Repeat Containing protein (BTRC) (Semplici et al., 2002).

References:

Semplici F, Meggio F, Pinna LA, Oliviero S (2002) CK2-dependent phosphorylation of the E2 ubiquitin conjugating enzyme UBC3B induces its interaction with beta-TrCP and enhances beta-catenin degradation. Oncogene 21, 3978-87.