Enabling Protein Degradation Drug Discovery

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  • Name
    Catalogue Number
    Size
    Price
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  • Name:
    UBE2Q1 (NICE-5) [6His-tagged]
    Catalogue Number:
    62-0081-100
    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
    ~50 kDa
  • Stability
    12 months at -70°C; aliquot as required
  • Protein Sequence
    Accession number: NP_060052. 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-UBE2Q1 was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the His-UBE2Q1 E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and His-UBE2Q1 enzymes in the presence of ubiquitin and ATP at 30°C was compared at two time points, T0 and T10 minutes.  The sensitivity of this ubiquitin/His-UBE2Q1 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). UBE2Q1 is a member of the E2 conjugating enzyme family and cloning of the human gene was described by Marenholz et al. (2001). UBE2Q1 shares 50-75% sequence identity with its homologues in Mus musculus, Drosophila, C. elegans and Xenopus. Murine UBE2Q1 has a conserved sequence for ubiquitin binding shared by all the ubiquitin-conjugating enzymes, however its NH(2)-terminal domain appears critical for the binding and internalization of cell surface galactosyltransferase 1 (GalT1) in embryonic stem cells. UBE2Q1 regulates GalT1-associated, laminin-dependent embryonic cell adhesion and the formation of embryoid bodies (Wassler et al., 2008).

References:

Marenholz I, Zirra M, Fischer DF, Backendorf C, Ziegler A, Mischke D (2001) Identification of human epidermal differentiation complex (EDC)-encoded genes by subtractive hybridization of entire YACs to a gridded keratinocyte cDNA library. Genome Res 11, 341-55.

Wassler MJ, Shur BD, Zhou W, Geng YJ (2008) Characterization of a novel ubiquitin-conjugating enzyme that regulates beta1,4-galactosyltransferase-1 in embryonic stem cells. Stem Cells 26, 2006-18.