Verified Applications: Recombinant Ubiquitin Conjugating Enzyme E2 2V1/2N accepts activated ubiquitin from Ubiquitin Activating Enzyme 1 (an E1) in in vitro reactions. This charged E2 may subsequently transfer ubiquitin to a protein substrate in an E3 Ligase-catalyzed reaction. Appropriate enzyme concentrations are specific to the application.
2 μg UBE2V1/UBE2N run on 4-12% SDS-PAGE gel under reducing conditions, then visualized with Colloidal Coomassie Blue Stain.
Formulation: 40 mM HEPES, 100 mM NaCl, 10% Glycerol, 1 mM EDTA, 1 mM TCEP, pH 7.6
Shipping: The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage and Stability: Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Aliquot and store ≤ -70°C (stable for 24 months from date of receipt).
Protein Sequence: GPGSMAATTGSGVKVPRNFRLLEELEEGQKGVGDGTVSWGLEDDEDMTLTRWTGMIIGPPRTIYENRIYSLKIECGPKYPEAPPFVRFVTKINMNGVNSSNGVVDPRAISVLAKWQNSYSIKVVLQELRRLMMSKENMKLPQPPEGQCYSN;GPGSMAGLPRRIIKETQRLLAEPVPGIKAEPDESNARYFHVVIAGPQDSPFEGGTFKLELFLPEEYPMAAPKVRFMTKIYHPNVDKLGRICLDILKDKWSPALQIRTVLLSIQALLSAPNPDDPLANDVAEQWKTNEAQAIETARAWTRLYAMNNI
Background Information and Alternate Names
Species & domain architecture
UBE2N is a 152-aa UBC fold with catalytic Cys87; UBE2V1 is 154 aa, conserving the β-sheet/α-helical scaffold but replacing the active cysteine with Ser32. Crystal and NMR structures of UBE2N~Ub bound to UBE2V1 (and yeast Mms2) reveal how UEV positions the acceptor ubiquitin (Ub) so that Lys63 lines up with UBE2N’s thioester.
UPS pathway role
The heterodimer works with diverse RING E3 enzymes. In innate-immunity, UBE2V1/UBE2N partners with TRAF6 to assemble Lys63 chains that recruit TAB2/TAK1 and activate NF-κB signaling. In genome maintenance, it collaborates with RAD5-like E3s to poly-ubiquitylate PCNA, licensing error-free template switching and fork reversal after DNA damage.
Lys63 chains orchestrate signaling, not degradation; mis-regulation underpins chronic inflammation, cancer metastasis and chemoresistance. Pharmacologic blockade of UBE2N dampens inflammatory cytokine output and sensitizes tumors to genotoxic drugs, making the complex a therapeutic target and a liability screen in degrader campaigns. UBE2N is believed to be a Lys63 “chain-building machine,” summarizing its emerging drugability.
References
Deng, L., et al., (2000) Cell 103:351-361. PMID 11057907
McKenna, S., et al., (2003) J Biol Chem 278:13151-13158. PMID 12569095
Eddins, M. J., et al., (2006) Nat. Struct. Mol. Biol. 13:915-920. PMID 16980971
Hodge, C.D., et al., (2016) Oncotarget 7:64471-64504. PMID 27486774
Vujanovic, M., et al., (2017) Mol Cell 67:882-890. PMID 28886337
