UBE2C (UbcH10) [6His-tagged]


Catalogue Number
62-0005-020
Product Size
20 µg
Price £
£100
Accession Number
NP_008950.1
Residues Expressed
1-179
Alternate Product Size
100 µg
Certificate of Analysis Size
20 µg
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
~23 kDa
Stability
12 months at -70°C; aliquot as required
Protein Sequence
Accession number: NP_008950.1. 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-UBE2C was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the His-UBE2C E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and His-UBE2C 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-UBE2C 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 regulated and targeted proteosomal 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). UBE2C is a member of the E2 ubiquitin-conjugating enzyme family and cloning of the human gene was first described by Townsley et al. (1997). UBE2C shares 85% and 61% homology with frog and clam sequences respectively, and contains a 30 amino acid N-terminal extension. UBE2C binds APC11 and APC2 resulting in their autoubiquitylation and proteosomal degradation, a switch in the APC complex which causes sister chromatid separation and exit from mitosis (Rape and Kirschner, 2004; Tang et al., 2001). UBE2C has also been identified as a molecular marker useful in non small cell lung carcinoma (NSCLC) prognosis (Kadara et al., 2009).


References

Kadara H, Lacroix L, Behrens C, Solis L, Gu X, Lee JJ, Tahara E, Lotan D, Hong WK, Wistuba II, Lotan R (2009) Identification of gene signatures and molecular markers for human lung cancer prognosis using an in vitro lung carcinogenesis system. Cancer Prev Res (Phila Pa) 2, 702-11.

Rape M, Kirschner MW (2004) Autonomous regulation of the anaphase-promoting complex couples mitosis to S-phase entry. Nature 432, 588-95.

Tang Z, Li B, Bharadwaj R, Zhu H, Ozkan E, Hakala K, Deisenhofer J, Yu H (2001) APC2 Cullin protein and APC11 RING protein comprise the minimal ubiquitin ligase module of the anaphase-promoting complex. Mol Biol Cell 12, 3839-51.

Townsley FM, Aristarkhov A, Beck S, Hershko A, Ruderman JV (1997) Dominant-negative cyclin-selective ubiquitin carrier protein E2-C/UbcH10 blocks cells in metaphase. Proc Natl Acad Sci USA 94, 2362-7.