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NameCatalogue NumberSizePriceAdd to Basket
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Catalogue Number:63-2004-025Size:25 µgPrice:£100Add To Basket
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Specieshuman
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SourceE. coli
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Quantity25µg
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Storage-70°C
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Concentration0.5 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 Weight33.3kDa
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Stability12 months at -70°C; aliquot as required
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Protein SequenceAccession number: NP_001014305.1. 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; ActivityE3 Ligase Assay: The activity of His-DCNL2 was validated through its ability to enhance the neddylation of Cul1/Rbx1 acting as a substrate in the presence of the thioester-loaded His-Ube2M~NEDD8. Incubation of Cul1/ Rbx1 and thioester loaded His-Ube2M~NEDD8 in thepresence or absence of His-DCNL2 at 4°C was compared at two time points T0 and T2 minutes. Increased neddylation of the Cul1 subunit in the presence of His-DCNL2 was demonstrated.
The enzymes of the NEDDylation pathway play a pivotal role in the activation of the largest class of ubiquitin E3 ligases called Cullin-RING-Ligases (CRLs). Akin to ubiquitylation three classes of enzymes are involved in the process of mammalian NEDDylation; E1 activating enzyme (APP-BP1/ UBA3 heterodimer), E2 conjugating enzymes (UBE2M or UBE2F) and E3 ligases the defective in Cul neddylation 1 domain-containing proteins (DCUN1D1-5) (Meyer-Schaller et al., 2009; Huang et al., 2011). There are 5 human DCUN1D1-5 proteins are also named defective in Cul neddylation 1 like proteins (DCNL1–5) (Meyer-Schaller et al., 2009). Cloning of DCNL2 was first described by Kurz et al. (2005) and Lamesch et al. (2007). The DCNLs have distinct amino-terminal domains, but share a conserved C-terminal potentiating neddylation (PONY) domain (Kurz et al., 2008). It has been determined that the interaction between the DCNLs and Cul1 occurs through the PONY domain and the Winged Helix DNA binding domain (WHB) respectively (Kurz et al., 2008; Scott et al., 2011). Pairwise analysis of 30 combinations of the five DCNLPONY domains and six cullin WHB subdomains by isothermal titration calorimetry have all shown interaction albeit with differing affinities (Monda et al., 2013).
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