UBE2R2 (CDC34B) [6His-tagged]


Catalogue Number
62-0094-100
Product Size
100 µg
Price £
£430
Accession Number
AAH04862
Residues Expressed
1-238
Alternate Product Size
20 µg
Certificate of Analysis Size
100 µg
Species
Human
Source
E. coli expression
Quantity
100 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.


Background

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.