2 μg UBE3C run on 4-12% SDS-PAGE gel under reducing conditions, then visualized with Colloidal Coomassie Blue Stain.

Active, Recombinant Human UBE3C

E3 ligases

Product Code: TE3-066

Verified Applications: In vitro, recombinant UBE3C accepts activated ubiquitin from E2 conjugating enzymes UBE2D1 or UBE2L3, in a reaction that also requires Ubiquitin Activating Enzyme 1 (an E1). The charged E3 demonstrates strong autoubiquitylation in the absence of added substrates. Appropriate enzyme concentrations are specific to the application.

Activity: Verified in Polyubiquitin Chain Synthesis Assay.

Purity: 95 %

Concentration: 50 – 70 μM

For Research Use Only (RUO)

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Note: To maintain product quality, larger quantities may be packaged as multiple smaller aliquots. The 1 mg size is generally supplied in a single vial.

UniProt #: Q15386

Predicted Molecular Weight: 58 kDa

Tag: N-6xHis-3C-SUMO

Background and Additional Info

Background: Ubiquitin-protein ligase E3C (UBE3C) is one of 28 HECT (Homologous to E6AP C-Terminus) class ubiquitin ligases found in humans. UBE3C’s HECT domain is an L-shaped, bi-lobed structure, with a large N-lobe and a small C-lobe. A region N-terminal to the HECT domain affect the stability and activity of UBE3C. Utilizing the E2 enzyme UBE2L3 or UBE2D1, UBE3C polyubiquitylates substrates, favoring K29 and K48 linkages. Reported substrates of the ligase include ADRM1, PIK3C3, CAND2, IRF3 and others. Anchored to 26S proteasomes, UBE3C elongates or regenerates ubiquitin chains on stalled substrates, facilitating their extraction.

Shipping: The product is shipped with dry ice. 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).

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References: Kane, E., et al. (2022) Biosci Rep 42:BSR20221036. PMID 36111624

Chu, B. W., et al., (2013) J Biol Chem 288:34575-87. PMID 24158444

Singh, S., and J. Sivaraman (2020) Biochem J 477:905-23. PMID 32039437

Hang, C., et al., (2021) Cancer Cell Int 21:25. PMID 33407510

Cui, B., et al., (2024) J Virol 98:e0133524. PMID 39212385

Alternate Names: HECTH2, RAUL, Ubiquitin-protein ligase E3C

Protein Sequence: MGGSHHHHHHGSLEVLFQGPGSMSDQEAKPSTEDLGDKKEGEYIKLKVIGQDSSEIHFKVKMTTHLKKLKESYCQRQGVPMNSLRFLFEGQRIADNHTPKELGMEEEDVIEVYQEQTGGVPFEERVKIFQRLIYADKQEVQGDGPFLDGINVTIRRNYIYEDAYDKLSPENEPDLKKRIRVHLLNAHGLDEAGIDGGGIFREFLNELLKSGFNPNQGFFKTTNEGLLYPNPAAQMLVGDSFARHYYFLGRMLGKALYENMLVELPFAGFFLSKLLGTSADVDIHHLASLDPEVYKNLLFLKSYEDDVEELGLNFTVVNNDLGEAQVVELKFGGKDIPVTSANRIAYIHLVADYRLNRQIRQHCLAFRQGLANVVSLEWLRMFDQQEIQVLISGAQVPISLEDLKSFTNYSGGYSADHPVIKVFWRVVEGFTDEEKRKLLKFVTSCSRPPLLGFKELYPAFCIHNGGSDLERLPTASTCMNLLKLPEFYDETLLRSKLLYAIECAAGFELS