2 μg AREL1 run on 4-12% SDS-PAGE gel under reducing conditions, then visualized with Colloidal Coomassie Blue Stain.

Active, Human, Recombinant AREL1

E3 ligases

Product Code: TE3-065

Verified Applications: In vitro, recombinant Apoptosis-resistant E3 ubiquitin-protein ligase 1 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 Poly-ubiquitin Chain Synthesis Assay.

Purity: 95%

Concentration: 20 – 30 μM

For Research Use Only (RUO)

Protein Name: AREL1

UniProt #: O15033

Predicted Molecular Weight: 59

Background and Additional Info

Background: Apoptosis-resistant E3 ubiquitin protein ligase 1 (AREL1) is one of 28 HECT (Homologous to E6AP C-Terminus) class ubiquitin ligases found in humans. AREL1 contains an N-terminal Filamin-like repeat, two WW motifs for substrate docking and a C-terminal HECT catalytic module (residues 436-823) with the active-site Cys790. High-resolution crystal data reveal a classic bilobed HECT fold capped by a unique β-hairpin that precisely positions the acceptor ubiquitin. Utilizing the E2 enzyme UBE2L3, AREL1 poly-ubiquitylates substrates, favoring K11 and K33 linkages. Evidence suggests AREL1-mediated ubiquitylation and degradation of cytosolic forms of IAP antagonists SMAC, HtrA2, and ARTS plays an important role in the regulation of apoptosis.

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).

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Additional Product Information

References:

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

Kim, J-B., et al. (2013) J Biol Chem 288(17):12014-21. PMID 23479728

Lear, T. B., et al., (2019) JCI Insight 4:e129110. PMID 31578312

Singh, S., et al. (2019) J Biol Chem 294:19934-19949. PMID 31732561

Jo, Y., et al. (2021) Exp Ther Med 22:1195. PMID 34584540

Roberts, J. Z., et al., (2022) Cell Death Differ. 29:272-284. PMID 34912054

Alternate Names: AREL1 (gene AREL1; synonyms KIAA0317, FLR)

Protein Sequence: MGWSHPQFEKGSHHHHHHGSLEVLFQGPGSMSDQEAKPSTEDLGDKKEGEYIKLKVIGQDSSEIHFKVKMTTHLKKLKESYCQRQGVPMNSLRFLFEGQRIADNHTPKELGMEEEDVIEVYQEQTGGSETFQDKVNFFQRELRQVHMKRPHSKVTLKVSRHALLESSLKATRNFSISDWSKNFEVVFQDEEALDWGGPRREWFELICKALFDTTNQLFTRFSDNNQALVHPNPNRPAHLRLKMYEFAGRLVGKCLYESSLGGAYKQLVRARFTRSFLAQIIGLRMHYKYFETDDPEFYKSKVCFILNNDMSEMELVFAEEKYNKSGQLDKVVELMTGGAQTPVTNANKIFYLNLLAQYRLASQVKEEVEHFLKGLNELVPENLLAIFDENELELLMCGTGDISVSDFKAHAVVVGGSWHFREKVMRWFWTVVSSLTQEELARLLQFTTGSSQLPPGGFAALCPSFQIIAAPTHSTLPTAHTCFNQLCLPTYDSYEEVHRMLQLAISEGCEGFGML