E-300

Material Data Sheet

GST-Ubiquitin E1 Enzyme (UBE1)

Price: $215.00
Catalog #: E-300

GST-Ubiquitin E1 Enzyme (UBE1)

Price: 
$215
Quantity: 
50 µg
Data Sheet: 

Ubiquitin­-activating Enzyme (UBE1), also known as Ubiquitin­-like Modifier Activating Enzyme 1 (UBA1), is a 1024 amino acid (aa) canonical member of the Ubiquitin-­activating (E1) enzyme family of proteins with a predicted molecular weight of 114 kDa.  Human and mouse UBE1 share 52% and 53% aa sequence identity with the yeast UBE1 protein, respectively. UBE1 is found in the cytoplasm and nucleus, and contains a conserved active­site cysteine residue and ATP-­binding site common to E1 enzymes. UBE1 is responsible for the first step in Ubiquitin­-protein isopeptide bond formation. Ubiquitin is activated by UBE1 and thereafter linked to the side chain of a cysteine residue in UBE1, Cys600 in S. cerevisiae, yielding a Ubiquitin-­UBE1 conjugate via a thioester bond.  The activated Ubiquitin is then transferred to a lysine residue on the target protein via the Ubiquitin­-conjugating – Ubiquitin ligase enzyme cascade. UBE1 is required for cell cycle progression and has been linked to cellular responses to DNA damage such as nucleotide excision repair.  Mutations in UBE1 are associated with X-­linked lethal infantile spinal muscular atrophy.  UBE1 is a critical component for the initiation of in vitro ubiquitin conjugation reactions.  This protein has an N­-terminal GST tag.

Product Information

Stock: 
0.71 mg/ml (5 µM) in 50 mM HEPES pH 8.0, 100 mM NaCl, 10% Glycerol (v/v), 1 mM TCEP
MW: 
141 kDa
Purity: 
> 90% by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie Blue stain.

Use & Storage

Use: 

Reaction conditions will need to be optimized for each specific application.  We recommend an initial GST-Ubiquitin E1 Enzyme concentration of 50-200 nM.

Storage: 

Store at -80°C and avoid multiple freeze-thaw cycles for best performance.

Literature

References: 

Haas, A.L. et al. (1982) J. Biol. Chem. 257:2543

Haas, A.L. & I.A. Rose (1982) J. Biol. Chem. 257:10329

Handley, P.M. et al. (1991) Proc. Natl. Acad. Sci. USA 88:258

Hershko, A. et al. (1983) J. Biol. Chem. 258:8206

Moundry, P. et al. (2012) Cell Cycle 11:1573

Nagai, Y. et al. (1995) J. Cell Sci. 108:2145

Nouspikel, T. & P.C. Hanawalt (2006) Proc. Natl. Acad. Sci. USA 103:16188

Schulman, B.A. & J.W. Harper (2009) Nat. Rev. Mol. Cell Biol. 10:319

Stephen, A.G. et al. (1996) J. Biol. Chem. 271:15608