Verified Applications: Recombinant Ubiquitin Conjugating Enzyme E2 D1 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 UBE2D1 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: GPGSMALKRIQKELSDLQRDPPAHCSAGPVGDDLFHWQATIMGPPDSAYQGGVFFLTVHFPTDYPFKPPKIAFTTKIYHPNINSNGSICLDILRSQWSPALTVSKVLLSICSLLCDPNPDDPLVPDIAQIYKSDKEKYNRHAREWTQKYAM
Background Information and Alternate Names
Species & domain architecture
The 152-aa protein consists of a compact UBC fold with the catalytic Cys85 on a protruding loop. Crystal structures of UbcH5a alone and in complex with ubiquitin (Ub) or RING domains show a rigid β-sheet core flanked by flexible loops that dictate linkage bias. No additional regulatory extensions are present, making it an ideal minimal E2 scaffold.
UPS pathway role
UBE2D1 forms thioester-linked UBE2D1~Ub intermediates that cooperate with diverse E3s—including CHIP, MDM2, RNF4, SMURF2, and Parkin—to ubiquitylate substrates involved in proteostasis, DNA repair, innate immunity, and mitophagy. Its ability to seed or elongate multiple chain types enables RING ligases to switch linkage patterns dynamically.
Relevance to TPD & disease biology
Over-expression of UBE2D1 correlates with enhanced migration and poorer prognosis in several cancers (e.g., gastric carcinoma), while its engagement by SMURF2 stabilizes oncogenic KRAS in lung and pancreatic tumors. Conversely, selective inhibition or knock-down can dampen NF-κB or Wnt signaling. Because most PROTAC workflows harness RING E3s that naturally recruit UBE2D1, the enzyme is central to optimizing degrader efficiency and minimizing off-pathway poly-ubiquitylation.
References
Xu, Z., et al., (2008) BMC Struct Biol 8:26. PMID 18485199
Bosanac, I., et al., (2011) J Mol Biol 408:420-431. PMID 21396940
Plechanovova, A., et al., (2012) Nature 489:115-120. PMID 22842904
Shukla, S., et al., (2014) Neoplasia 16:115-128. PMID 24709419
