Active Recombinant Human CUL1/RBX1/SKP1

• Cat.No.: CUL1-147H
• Product Overview: Recombinant human CUL1 (amino acid residues 1-776), RBX1 (amino acid residues 1-108), SKP1 (amino acid residues 1-160) was expressed in Insect (Sf21) cells.

Specification

• Species: Human
• Source: Insect Cells
• Tag: Non
• Protein Length: 1-776;1-108;1-160 a.a.
• Description: Cullin-RING-Ligases (CRLs) are one largest class of ubiquitin E3 ligases and the enzymes of the NEDDylation pathway play a pivotal role in the activation of these, akin to ubiquitylation, the E1 activating enzyme (APP-BP1/UBA3 heterodimer) and the E2 conjugating enzymes (UBE2M or UBE2F) are involved in mammalian NEDDylation of the Cullin Ring Ligases (CRLs) (Meyer-Schaller et al., 2009; Huang et al., 2011; Morimoto et al., 2003).
• Form: 50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol
• Bio-activity: E3 Ligase Assay: The activity of Cul1/Rbx1/Skp1 was validated indirectly through its ability to act as a substrate for Neddylation in the presence of the NEDD8 E3 ligase GST-DCNL1 and thioester-loaded His-Ube2M-NEDD8. Incubation of Cul1/Rbx1/Skp1 and thioester loaded His-Ube2M-NEDD8 in the presence or absence of GST-DCNL1 at 4?C was compared at two time points T0 and T2 minutes. Increased Neddylation of the Cul1 subunit in the presence of GST-DCNL1 was demonstrated.
• Molecular Mass: Cul1: ~89.9 kDa; Rbx1: ~12.3 kDa; Skp1: ~18.3
• Purity: >95% by InstantBlue™ SDS-PAGE
• Storage: 12 months at -70°C. Avoid multiple freeze/thaw cycles.
• Concentration: 0.5 mg/ml
• Publications:
  Lys29-linkage of ASK1 by Skp1? Cullin 1? Fbxo21 ubiquitin ligase complex is required for antiviral innate response." (2016)

Gene Information

• Gene Name: CUL1 cullin 1 [ Homo sapiens ]
• Official Symbol: CUL1
• Synonyms: CUL1; cullin 1; cullin-1; CUL-1; MGC149834; MGC149835
• Gene ID: 8454
• mRNA Refseq: NM_003592
• Protein Refseq: NP_003583
• MIM: 603134
• UniProt ID: Q13616
• Chromosome Location: 7q36.1
• Pathway: APC/C-mediated degradation of cell cycle proteins, organism-specific biosystem; Activation of NF-kappaB in B Cells, organism-specific biosystem; Adaptive Immune System, organism-specific biosystem; Antigen processing: Ubiquitination and Proteasome degradation, organism-specific biosystem; Canonical Wnt signaling pathway, organism-specific biosystem; Cell Cycle, organism-specific biosystem
• Function: protein binding; ubiquitin protein ligase binding

Citation

Lys29-linkage of ASK1 by Skp1?Cullin 1?Fbxo21 ubiquitin ligase complex is required for antiviral innate response

Journal: eLife    Data: 2016/4/11

Authors: Zhou Yu, Taoyong Chen, Jianli Wang

Article Snippet:The in vitro analysis of ubiquitination of ASK1 was performed as described (Chen et al., 2013) with modifications.The in vitro analysis of ubiquitination of ASK1 was performed as described (Chen et al., 2013) with modifications.. The ubiquitination of ASK1 was performed in 50 ml buffer containing 40 mM TrisHCl, pH 7.1, 40 mM NaCl, 1 mM b-ME, 5 mM MgCl2 supplemented with 2 mM ATP, 1 mM ubiquitin aldehyde, 1 mg Conjugation Fraction A (F-340, BostonBiochem, Cambridge, MA), 1 mg/ml ubiquitin and derivatives (BostonBiochem), 10 ng/ml recombinant ASK1 (MAP3K5-9515M) and Cul1-Rbx1Skp1 (CUL1-147H) (Creative BioMart, Shirley, NY) and 10 ng/ml GST-Fbxo21 in the presence or absence of Nedd8 Conjugation System (#J3150, UBPBio, Aurora, CO) and 5 mM MLN-4924 for 30 min at 37?C.. Reaction products were finally processed for Ub immunoblotting.Reaction products were finally processed for Ub immunoblotting.