UBE2H (UbcH2) [6His-tagged]

Ubiquigent

Catalog No: UBI-62-0078-020

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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_003335. 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-UBE2H  was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the His-UBE2H  E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and His-UBE2H  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-UBE2H  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 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). UBE2H is a member of the E2 conjugating enzyme family and cloning of the human gene was first described by Kaiser et al. (1994). Human UBE2H shares 54% identity with its yeast homologue (UbcH8) and full length forms of both the human and yeast enzymes showed similar enzymatic activities in vitro by catalyzing the ubiquitylation of histones (Kaiser et al., 1995). In skeletal muscle whole cell extracts TNFα up-regulates and increases the conjugating activity of UBE2H in vitro via binding of NFκB to the promoter region of the gene (Li et al., 2003). UBE2H has been mapped to a region on chromosome 7 (Hayashida et al., 2000) and the gene has been identified as a candidate for involvement in an autistic disorder with neurodevelopmental complications. Single strand conformation analysis demonstrated a significant association between a polymorphism in the UBE2H gene suggesting it could be one of the 7q-susceptibility loci for Autistic Disorder (Vourc’h et al., 2003). An association has also been made between UBE2H and the motor neuron disorder amyotrophic lateral sclerosis (ALS) where single strand conformation polymorphism (SSCP) analysis identified a known and sporadic polymorphism in the gene (Martin et al., 2008).

References:

Hayashida S, Yamasaki K, Asada Y, Soeda E, Niikawa N, Kishino T (2000) Construction of a physical and transcript map flanking the imprinted MEST/PEG1 region at 7q32. Genomics 66, 221-5.
Kaiser P, Mandl S, Schweiger M, Schneider R (1995) Characterization of functionally independent domains in the human ubiquitin conjugating enzyme UbcH2. FEBS Lett 377, 193-6.
Kaiser P, Seufert W, Hofferer L, Kofler B, Sachsenmaier C, Herzog H, Jentsch S, Schweiger M, Schneider R (1994) A human ubiquitin-conjugating enzyme homologous to yeast UBC8. J Biol Chem 269, 8797-802.
Li YP, Lecker SH, Chen Y, Waddell ID, Goldberg AL, Reid MB (2003) TNF-alpha increases ubiquitin-conjugating activity in skeletal muscle by up-regulating UbcH2/E220k. FASEB J 17, 1048-57.
Martin I, Vourc’h P, et al. (2009) Association study of the ubiquitin conjugating enzyme gene UBE2H in sporadic ALS. Amyotroph Lateral Scler 10, 432-5.
Vourc’h P, Martin I, Bonnet-Brilhault F, Marouillat S, Barthelemy C, Pierre Muh J, Andres C (2003) Mutation screening and association study of the UBE2H gene on chromosome 7q32 in autistic disorder. Psychiatr Genet 13, 221-5.