UBE2J1 (NCUBE1) residues (1-282) [6His-tagged]

Ubiquigent

Catalog No: UBI-62-0096-020

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Species Reactivity: Human  
Source: E. coli expression  
Formula: 50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol  
Concentration: 1 mg/ml  
Molecular Weight: ~35 kDa  
Protein Sequence: Accession number: NP_057105.2. For full protein sequence information download the Certificate of Analysis pdf.  
Quality Control Protein Identification: Confirmed by mass spectrometry.  
Quality Control Activity: E2-Ubiquitin Thioester Loading Assay: The activity of His-UBE2J1 was validated by loading E1 UBE1 activated ubiquitin onto the active cysteine of the His-UBE2J1 E2 enzyme via a transthiolation reaction. Incubation of the UBE1 and His-UBE2J1 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/His-UBE2J1 thioester bond to the reducing agent DTT was confirmed.   

Background

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). UBE2J1 is a member of the E2 conjugating enzyme family and cloning of the human gene was first described by Lester et al. (2000). UBE2J1 is a 318 amino acid single-pass type IV membrane protein, which can be found localised to the membrane of the endoplasmic reticulum (ER) (Lester et al., 2000). UBE2J1 catalyzes the modification of misfolded membrane proteins with ubiquitin which results in their targeting to the proteasome for degradation (Walter et al., 2001). UBE2J1 has been shown to interact with the E3 ligase ICP0 forming polyubiquitin chains in an in vitro polyubiquitylation assay. ICP0 targets substrates such as PML, Sp100, CENP-C, and CENP-A for proteasomal degradation (Everett et al., 1999; Everett et al., 1998; Lomonte et al., 2001; Parkinson and Everett. 2000). Evidence for an association of the UBE2J1 gene locus with Serum creatinine (S CR) levels has been demonstrated. S CR is a biomarker used for the non-invasive assessment of kidney function and it is hoped will provide an insight into the genetic basis of serum creatinine regulatory processes (Pattaro et al., 2010).

References:

Everett RD, Earnshaw WC, Findlay J, Lomonte P (1999) Specific destruction of kinetochore protein CENP-C and disruption of cell division by herpes simplex virus immediate-early protein Vmw110. EMBO J 18, 1526-38.
Everett RD, Freemont P, Saitoh H, Dasso M, Orr A, Kathoria M, Parkinson J (1998) The disruption of ND10 during herpes simplex virus infection correlates with the Vmw110- and proteasome-dependent loss of several PML isoforms. J Virol 72, 6581-91.
Lester D, Farquharson C, Russell G, Houston B (2000) Identification of a family of noncanonical ubiquitin-conjugating enzymes structurally related to yeast UBC6. Biochem Biophys Res Commun 269, 474-80.
Lomonte P, Sullivan KF, Everett RD (2001) Degradation of nucleosome-associated centromeric histone H3-like protein CENP-A induced by herpes simplex virus type 1 protein ICP0. J Biol Chem 276, 5829-35.
Parkinson J, Everett RD (2000) Alphaherpesvirus proteins related to herpes simplex virus type 1 ICP0 affect cellular structures and proteins. J Virol 74, 10006-17.
Pattaro C, De Grandi A, et al.(2010) A meta-analysis of genome-wide data from five European isolates reveals an association of COL22A1, SYT1, and GABRR2 with serum creatinine level. BMC Med Genet 11, 41.
Walter J, Urban J, Volkwein C, Sommer T (2001) Sec61p-independent degradation of the tail-anchored ER membrane protein Ubc6p. EMBO J 20, 3124-31.