Recombinant Yeast GST-Ubiquitin Activating Enzyme (UBE1), CF

R&D Systems | Catalog # E-300

Will be discontinued when existing inventory is gone.
R&D Systems

Key Product Details

Source

Sf 21 (baculovirus)

Accession Number

P22515.2

Applications

Bioactivity
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Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived yeast Ubiquitin-activating Enzyme/UBE1 protein
Met1 - Leu1024 with a N-terminal GST (glutathione S-transferase) tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Predicted Molecular Mass

141 kDa

Activity

Recombinant Yeast GST-Ubiquitin E1 Enzyme (UBE1) is a member of the Ubiquitin-activating (E1) enzyme family that is required for the first step of the enzymatic cascade that subsequently utilizes a Ubiquitin-conjugating (E2) enzyme and a Ubiquitin ligase (E3) to conjugate Ubiquitin to substrate proteins.  Reaction conditions will need to be optimized for each specific application.   We recommend an initial GST-Ubiquitin E1 Enzyme (UBE1) concentration of 50-200 nM.

Formulation, Preparation, and Storage

E-300
Formulation Supplied as a solution in HEPES, NaCl, TCEP and Glycerol.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Ubiquitin-activating Enzyme/UBE1

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 active site cysteine residue and ATP-binding site common to E1 enzymes (1-3).  UBE1 is responsible for the first step in Ubiquitin-protein isopeptide bond formation (4,5).  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 (5-8).  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 (3,9,10). Mutations in UBE1 are associated with X-linked lethal infantile spinal muscular atrophy (11).  UBE1 is a critical component for the initiation of in vitro ubiquitin conjugation reactions.  

References

  1. Handley, P.M. et al. (1991) Proc. Natl. Acad. Sci. USA 88:258.
  2. Nagai, Y. et al. (1995) J. Cell Sci. 108:2145.
  3. Stephen, A.G. et al. (1996) J. Biol. Chem. 271:15608.
  4. Hershko, A. et al. (1983) J. Biol. Chem. 258:8206.
  5. Schulman, B.A. and J.W. Harper (2009) Nat. Rev. Mol. Cell Biol. 10:319.
  6. Haas, A.L. et al. (1982) J. Biol. Chem. 257:2543.
  7. Haas, A.L. and I.A. Rose (1982) J. Biol. Chem. 257:10329.
  8. Pickart, C.M. et al. (1994) J. Biol. Chem. 269:7115.
  9. Nouspikel, T. and P.C. Hanawalt (2006) Proc. Natl. Acad. Sci. USA 103:16188.
  10. Moundry, P. et al. (2012) Cell Cycle 11:1573.
  11. Ramser, J. et al. (2008) Am. J. Hum. Genet. 82:188.

Alternate Names

A1S9T, AMCX1, GXP1, POC20, SMAX2, UBA1, UBE1, Ubiquitinactivating Enzyme

Entrez Gene IDs

7317 (Human); 22201 (Mouse); 314432 (Rat); 100009225 (Rabbit); 853670 (Yeast)

Gene Symbol

UBA1

UniProt

P22515.2

Additional Ubiquitin-activating Enzyme/UBE1 Products

Product Documents for Recombinant Yeast GST-Ubiquitin Activating Enzyme (UBE1), CF

Certificate of Analysis

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Product Specific Notices for Recombinant Yeast GST-Ubiquitin Activating Enzyme (UBE1), CF

For research use only

Related Research Areas

E1 Ubiquitin/Ubl Activating Enzymes

Citations for Recombinant Yeast GST-Ubiquitin Activating Enzyme (UBE1), CF

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Associated Pathways

Ubiquitination Cascade Pathway Ubiquitination Cascade Pathway Thumbnail