Enabling Protein Degradation Drug Discovery

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  • Name
    Catalogue Number
    Size
    Price
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  • Name:
    UBE2J2 (NCUBE2) residues (1-226) [GST-tagged]
    Catalogue Number:
    62-0036-020
    Size:
    20 µg
    Price:
    £100
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  • Species
    Human
  • Source
    E. coli expression
  • Quantity
    20 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
    ~52 kDa
  • Stability
    12 months at -70°C; aliquot as required
  • Protein Sequence
    Accession number: NP_477515.2. 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 GST-UBE2J2 was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the GST-UBE2J2 E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and GST-UBE2J2 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-UBE2J2 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 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). UBE2J2 is a member of the E2 conjugating enzyme family and the human gene was first described by Lenk et al. (2002). UBE2J2 is a 318 amino acid protein that shares 26% identity to its yeast homologue (Lenk et al., 2002). UBE2J2 is localised to the cytoplasmic surface of the Endoplasmic Reticulum (ER) and participates in Endoplasmic Reticulum Associated Degradation (ERAD). It has been demonstrated that expression of a mutant form of UBE2J2 affects ERAD of the T cell receptor and a mutant form of the CFTR protein (Lenk et al., 2002). UBE2J2 has also been identified as the primary cellular E2 recruited by the E3 ligase murine K3 (mK3), and this E2-E3 pair is capable of conjugating ubiquitin on lysine or serine residues of substrates. Interestingly, UBE2J2-mK3 preferentially promotes ubiquitylation of hydroxylated amino acids via ester bonds even when lysine residues are present on wild-type substrates (Wang et al., 2009). Treatment of Sertoli and germ cells of the testies with a proteasome inhibitor results in the colocalisation of the retinoic acid receptor alpha (RARA) to the ER where it interacts with UBE2J2 resulting in its ubiquitylation and degradation via the ERAD pathway (Zhu et al., 2010).

References:

Lenk U, Yu H, Walter J, Gelman MS, Hartmann E, Kopito RR, Sommer T (2002) A role for mammalian Ubc6 homologues in ER-associated protein degradation. J Cell Sci 115, 3007-14.

Wang X, Herr RA, Rabelink M, Hoeben RC, Wiertz EJ, Hansen TH (2009) Ube2j2 ubiquitinates hydroxylated amino acids on ER-associated degradation substrates. J Cell Biol 187, 655-68.

Zhu L, Santos NC, Kim KH (2010) Disulfide isomerase glucose-regulated protein 58 is required for the nuclear localization and degradation of retinoic acid receptor alpha. Reproduction 139(4), 717-31.