UBE2Q2 [6His-tagged]


Catalogue Number
62-0051-100
Product Size
100 µg
Price £
£325
Accession Number
AAH34342.1
Residues Expressed
1-375
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
~45 kDa
Stability
12 months at -70°C; aliquot as required
Protein Sequence
Accession number: AAH34342.1. 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-UBE2Q2 was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the His-UBE2Q2 E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and His-UBE2Q2 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-UBE2Q2 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). UBE2Q2 is a member of the E2 conjugating enzyme family. The cloning of human UBE2Q2 was first described by Crawford and Piwnica-Worms. (2001). UBE2Q2 has been found to be up-regulated in 85% of head and neck squamous cell carcinoma tumours, with an increase of 2.4-fold compared to normal tissue. Immunohistochemistry and in situ hybridization analysis on tumour tissue sections has revealed strong signals in the tumour cell islets, invasive epithelia, and dysplastic regions (Seghatoleslam et al., 2006). UBE2Q2 has been identified as a novel oncosuppressor that inhibits tumour growth and it is thought it could function as a novel diagnostic tool and potential therapeutic target for head and neck squamous cell carcinoma (Maeda et al., 2009). UBE2Q2 may play a role in cytoskeleton structure and regulation, as actin and 6 actin-binding proteins have been shown to interact with UBE2Q2 (Seghatoleslam et al., 2006). Inhibition of UBE2Q2 following treatment of HeLa cells with Microtubule Inhibiting Agent (MIA) causes mitotic arrest and increased cytotoxicity, effects only observed in the absence of MIA suggesting UBE2Q2 is only involved in this response rather than having a more general role in mitosis (Banerjee et al., 2007).


References

Banerjee S, Brooks WS, Crawford DF (2007) Inactivation of the ubiquitin conjugating enzyme UBE2Q2 causes a prophase arrest and enhanced apoptosis in response to microtubule inhibiting agents. Oncogene 26, 6509-17.

Crawford DF, Piwnica-Worms H (2001) The G(2) DNA damage checkpoint delays expression of genes encoding mitotic regulators. J Biol Chem 276, 37166-77.

Maeda H, Miyajima N, Kano S, Tsukiyama T, Okumura F, Fukuda S, Hatakeyama S (2009) Ubiquitin-conjugating enzyme UBE2Q2 suppresses cell proliferation and is down-regulated in recurrent head and neck cancer. Mol Cancer Res 7, 1553-62.

Seghatoleslam A, Zambrano A, Millon R, Ganguli G, Argentini M, Cromer A, Abecassis J, Wasylyk B (2006) Analysis of a novel human gene, LOC92912, over-expressed in hypopharyngeal tumours. Biochem Biophys Res Commun 339, 422-9.