UBE2D4 (UbcH5d) [GST-tagged]

Ubiquigent

Catalog No: UBI-62-0016-020

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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: ~44 kDa  
Protein Sequence: Accession number: NP_057067. 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 GST-UBE2D4 was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the GST-UBE2D4 E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and GST-UBE2D4 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/GST-UBE2D4 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). UBE2D4 is a member of the E2 ubiquitin-conjugating enzyme family and the human gene was first described by Colland et al. (2004).

Reference:

Colland F, Jacq X, Trouplin V, Mougin C, Groizeleau C, Hamburger A, Meil A, Wojcik A, Legrain P, Gauthier J (2004) Functional proteomics mapping of a human signaling pathway. Genome Res 14, 1324-32.