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NameCatalogue NumberSizePriceAdd to Basket
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Catalogue Number:62-0025-100Size:100 µgPrice:£325Add To Basket
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SpeciesHuman
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SourceE. coli expression
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Quantity100 μg
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Storage-70°C
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Concentration1 mg/ml
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Formulation50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol
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Molecular Weight~23 kDa
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Stability12 months at -70°C; aliquot as required
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Protein SequenceAccession number: NP_542409. For full protein sequence information download the Certificate of Analysis pdf.
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QA; Protein IdentificationConfirmed by mass spectrometry.
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QA; ActivityE2-NEDD8 Thioester Loading Assay: The activity of UBE2F was validated by loading E1 (APP-BP1/Uba3) activated NEDD8 onto the active cysteine of the UBE2F E2 enzyme via a transthiolationreaction. Incubation of the APP-BP1/Uba3 and UBE2F enzymes in the presence of NEDD8 and ATP at 30°C was compared at two time points, T0 and T10 minutes. The sensitivity of the NEDD8/UBE2F thioester bond to the reducing agent DTT was confirmed.
The enzymes of the NEDDylation pathway play a pivotal role in a number of cellular processes including the indirect regulation and targeting of substrate proteins for proteasomal degradation. Three classes of enzymes are involved in the process of NEDDylation; the ubiquitin-like activating enzyme APP-BP1/Uba3 (E1), the ubiquitin-like conjugating enzymes (E2s) and protein ligases (E3s). UBE2F is a member of the E2 conjugating enzyme family and the human gene was first described by Huang et al. (2009). UBE2F acts as a NEDD8 conjugating enzyme both in vitro and in vivo. UBE2F accepts the ubiquitin-like protein NEDD8 from the Uba3-NAE1 (APP-BP1/Uba3) E1 complex and catalyzes its covalent attachment to other proteins. The specific interaction of UBE2F with the E3 ubiquitin ligase RBX2, but not RBX1, suggests that the RBX2-UBE2F complex NEDDylates specific target proteins such as CUL5, a component of one of the many Cullin Ring Ligases (CRLs) (Huang et al., 2009).
Reference:
Huang DT, Ayrault O, Hunt HW, Taherbhoy AM, Duda DM, Scott DC, Borg LA, Neale G, Murray PJ, Roussel MF, Schulman BA (2009) E2-RING expansion of the NEDD8 cascade confers specificity to cullin modification. Mol Cell 33, 483-95.