Enabling Protein Degradation Drug Discovery

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  • Name
    Catalogue Number
    Size
    Price
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  • Name:
    UBE2A (HR6A) [untagged]
    Catalogue Number:
    62-0002-100
    Size:
    100 µg
    Price:
    £325
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  • 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
    ~17 kDa
  • Stability
    12 months at -70°C; aliquot as required
  • Protein Sequence
    Accession number: NP_003327. 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 UBE2A was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the UBE2A E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and UBE2A 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/UBE2A thioester bond to the reducing agent DTT was confirmed.
     

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). UBE2A is a member of the E2 conjugating enzyme family and cloning of the human gene was first described by Koken et al. (1991). UBE2A shares 70% identity with its yeast homologue but lacks the acidic C-terminal domain. The ring finger proteins RAD5 and RAD18 interact with UBE2A and other members of the RAD6 pathway (Ulrich and Jentsch, 2000). Phosphorylation of UBE2A by CDK1 and 2 increases its activity during the G2/M phase of the cell cycle (Sarcevic et al., 2002). UBE2A is required for post-replicative DNA damage repair in eukaryotic cells and it is thought binding to ZNF198 may be involved in this process (Kunapuli et al., 2003). A nonsense mutation resulting in the loss of a 25 amino acid region in the C-terminal domain of UBE2A has been identified as a cause of a novel X-linked mental retardation (XLMR) syndrome (Nascimento et al., 2006).

References:

Koken MH, Reynolds P, Jaspers-Dekker I, Prakash L, Prakash S, Bootsma D, Hoeijmakers JH (1991) Structural and functional conservation of two human homologs of the yeast DNA repair gene RAD6. Proc Natl Acad Sci USA 88, 8865-9.

Kunapuli P, Somerville R, Still IH, Cowell JK (2003) ZNF198 protein, involved in rearrangement in myeloproliferative disease, forms complexes with the DNA repair-associated HHR6A/6B and RAD18 proteins. Oncogene 22, 3417-23.

Nascimento RM, Otto PA, de Brouwer AP, Vianna-Morgante AM (2006) UBE2A, which encodes a ubiquitin-conjugating enzyme, is mutated in a novel X-linked mental retardation syndrome. Am J Hum Genet 79, 549-55.

Sarcevic B, Mawson A, Baker RT, Sutherland RL (2002) Regulation of the ubiquitin-conjugating enzyme hHR6A by CDK-mediated phosphorylation. EMBO J 21, 2009-18.

Ulrich HD, Jentsch S (2000) Two RING finger proteins mediate cooperation between ubiquitin-conjugating enzymes in DNA repair. EMBO J 19, 3388-97.