2 μg UBA1 run on 4-12% SDS-PAGE gel under reducing conditions, then visualized with Colloidal Coomassie Blue Stain.

Active, Human, Recombinant UBA1, His-Strep II tag

E1 activating enzymes

Product Code: TE1-001

Verified Applications: Recombinant Ubiquitin Activating Enzyme 1 activates ubiquitin for subsequent transfer to a Ubiquitin Conjugating Enzyme (E2) in in vitro reactions. This initial event in the ubiquitin-proteasome cascade requires Mg-ATP and ubiquitin. Appropriate enzyme concentrations are specific to the application.

Activity: Verified in Ubiquitin Charging Assay. Verified in UbiREAL Assay.

Purity: 98%

Concentration: 5 μM

For Research Use Only (RUO)

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Protein Name: UBA1, tagged

UniProt #: P22314

Predicted Molecular Mass: 121 kDa

Background and Additional Info

Background: Ubiquitin-like modifier-activating enzyme 1 (UBA1, UBE1) is the primary enzyme (E1) responsible for the initial activation of ubiquitin, setting the stage for subsequent steps in ubiquitylation. UBA1 binds ATP and ubiquitin, then creates a high-energy ubiquitin-AMP intermediate while releasing pyrophosphate (PPi). Ubiquitin is then transferred to Cys632 as a thioester intermediate, releasing AMP. Finally, domain swiveling and an E2-docking Ubiquitin Fold Domain (UFD) ensure efficient transfer of the ubiquitin (transthiolation reaction) to a host of cellular E2 conjugating enzymes.

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. 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:

Olsen, S.K. and C. D. Lima (2013) Mol Cell 49:884-896. PMID 23416107

Schafer, A., et al., (2014) Acta Crystallogr D Biol. Crystallogr. 70:1311-1320. PMID 24816100

Groen, E. J. N., and T. H. Gillingwater (2015) Trends Mol Med 21:622-632. PMID

Barghout, S. H., et al., (2019) Leukemia 33:37-51. PMID 29884901

Poulter, J. A., et al., (2021) Blood 137:3676-3681. PMID 33690815

Alternate Names: UBE1, A1S9

Protein Sequence: MGWSHPQFEKGSHHHHHHHHGSLEVLFQGPGSMSSSPLSKKRRVSGPDPKPGSNCSPAQSVLSEVPSVPTNGMAKNGSEADIDEGLYSRQLYVLGHEAMKRLQTSSVLVSGLRGLGVEIAKNIILGGVKAVTLHDQGTAQWADLSSQFYLREEDIGKNRAEVSQPRLAELNSYVPVTAYTGPLVEDFLSGFQVVVLTNTPLEDQLRVGEFCHNRGIKLVVADTRGLFGQLFCDFGEEMILTDSNGEQPLSAMVSMVTKDNPGVVTCLDEARHGFESGDFVSFSEVQGMVELNGNQPMEIKVLGPYTFSICDTSNFSDYIRGGIVSQVKVPKKISFKSLVASLAEPDFVVTDFAKFSRPAQLHIGFQALHQFCAQHGRPPRPRNEEDAAELVALAQAVNARALPAVQQNNLDEDLIRKLAYVAAGDLAPINAFIGGLAAQEVMKACSGKFMPIMQWLYFDALECLPEDKEVLTEDKCLQRQNRYDGQVAVFGSDLQEKLGKQKYFLVGAGAIGCELLKNFAMIGLGCGEGGEIIVTDMDTIEKSNLNRQFLFRPWDVTKLKSDTAAAAVRQMNPHIRVTSHQNRVGPDTERIYDDDFFQNLDGVANALDNVDARMYMDRRCVYYRKPLLESGTLGTKGNVQVVIPFLTESYSSSQDPPEKSIPICTLKNFPNAIEHTLQWARDEFEGLFKQPAENVNQYLTDPKFVERTLRLAGTQPLEVLEAVQRSLVLQRPQTWADCVTWACHHWHTQYSNNIRQLLHNFPPDQLTSSGAPFWSGPKRCPHPLTFDVNNPLHLDYVMAAANLFAQTYGLTGSQDRAAVATFLQSVQVPEFTPKSGVKIHVSDQELQSANASVDDSRLEELKATLPSPDKLPGFKMYPIDFEKDDDSNFHMDFIVAASNLRAENYDIPSADRHKSKLIAGKIIPAIATTTAAVVGLVCLELYKVVQGHRQLDSYKNGFLNLALPFFGFSEPLAAPRHQYYNQEWTLWDRFEVQGLQPNGEEMTLKQFLDYFKTEHKLEITMLSQGVSMLYSFFMPAAKLKERLDQPMTEIVSRVSKRKLGRHVRALVLELCCNDESGEDVEVPYVRYTIR