UBE2C (UbcH10) [6His-tagged]

Ubiquigent

Catalog No: UBI-62-0005-020

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Species Reactivity: Human  
Source: E. coli expression  
Formula: 50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol  
Concentration: 1 mg/ml  
Molecular Weight: ~23 kDa  
Protein Sequence: Accession number: NP_008950.1. For full protein sequence information download the Certificate of Analysis pdf.  
Quality Control Protein Identification: Confirmed by mass spectrometry.  
Quality Control 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.