UBE1, Active, Recombinant, Human, 6-His-Tag (Ubiquitin activating enzyme E1, Ubiquitin-like modifier-activating enzyme 1, UBA1, A1S9 Protein)

UBE1 catalyzes the first step in ubiquitin conjugation to mark cellular proteins for degradation. It is responsible for the primary ubiquitin-protein isopeptide bond formation and is a critical component for the initiation of in vitro conjugation reactions. UBE1 activates ubiquitin by the ATP dependent adenylation of its C-terminal glycine carboxyl group. This ubiquitin residue is then linked to the sulphydryl side chain moiety of a cysteine residue within UBE1 by forming a high energy thiol ester bond, liberating free AMP. There are two active sites within the UBE1 molecule allowing it to accommodate two ubiquitin molecules at one time, with a new ubiquitin forming an adenylate intermediate as the previous one is transferred to the thiol site. The activated ubiquitin is then transferred to the lysine of target proteins via the E2/E3 conjugation cascade.\n\nSource:\nRecombinant corresponding to full length human UBE1, active, fused to 6His-tagged at N-terminus, expressed by baculovirus in Sf21 insect cells.\n\nMolecular Weight: \n~122kD\n\nStorage and Stability:\nAliquot to avoid repeated freezing and thawing and store at -70 degrees C. Aliquots are stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

029635

Size

10ug

Reactivities

Hum

Form

Supplied as a liquid in 50mM Tris/HCl pH7.5, 150mM sodium chloride, 0.1mM EGTA, 0.03% Brij-35, 270mM sucrose, 1mM benzamidine, 0.2mM PMSF, 0.1% 2-mercaptoethanol.

Purity

~94% (SDS-PAGE, Coomassie blue staining)

References

1. Haas A.L. and Rose I.A. The Mechanism of Ubiquitin Activating Enzyme. A Kinetic and Equilibrium Analysis. J Biol Chem., 257: 10329-10337, 1982 2. Haas A.L., et al. Ubiquitin-activating Enzyme. Mechanism and Role in protein Ubiquitin Conjugation. J Biol Chem., 257: 2543-2548, 1982 3. Handley P.M., et al. Molecular cloning, sequence, and tissue distribution of the human ubiquitin-activating enzyme E1. Proc. Nail. Acad. Sci. USA., 88: 258-262, 1991 4. Lee I, and Schindelin H., Structural Insights into E1-Catalyzed Ubiquitin Activation and Transfer to Conjugating Enzymes. Cell, 134: 268-278, 2008.