• Purity

  >98%, by HPLC

• Activity

  Add Recombinant Human Ubiquitin-Vinyl Sulfone to     in vitro assays to inhibit deubiquitinating enzymes. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human Ubiquitin-Vinyl Sulfone concentration of 1-5 μM.

• Source

  E. coli-derived Contains underivatized and C-terminal VS derivatized protein, quantity is by derivatized content

• Accession #

• Predicted Molecular Mass

  8.6 kDa

Background: Ubiquitin

Ubiquitin is a 76 amino acid (aa) protein that is ubiquitously expressed in all eukaryotic organisms. Ubiquitin is highly conserved with 96% aa sequence identity shared between human and yeast Ubiquitin, and 100% aa sequence identity shared between human and mouse Ubiquitin (1). In mammals, four Ubiquitin genes encode for two Ubiquitin-ribosomal fusion proteins and two poly-Ubiquitin proteins. Cleavage of the Ubiquitin precursors by deubiquitinating enzymes gives rise to identical Ubiquitin monomers each with a predicted molecular weight of 8.6 kDa. Conjugation of Ubiquitin to target proteins involves the formation of an isopeptide bond between the C-terminal glycine residue of Ubiquitin and a lysine residue in the target protein. This process of conjugation, referred to as ubiquitination or ubiquitylation, is a multi-step process that requires three enzymes: a Ubiquitin-activating (E1) enzyme, a Ubiquitin-conjugating (E2) enzyme, and a Ubiquitin ligase (E3). Ubiquitination is classically recognized as a mechanism to target proteins for degradation and as a result, Ubiquitin was originally named ATP-dependent Proteolysis Factor 1 (APF-1) (2,3). In addition to protein degradation, ubiquitination has been shown to mediate a variety of biological processes such as signal transduction, endocytosis, and post-endocytic sorting (4-7).

Ubiquitin-Vinyl Sulfone is a potent, irreversible and specific inhibitor of Ubiquitin C-terminal hydrolases (UCHs), Ubiquitin-specific proteases (USPs) and deubiquitinating enzymes (DUBs). This protein is useful for inhibiting the hydrolysis of poly-Ubiquitin chains on substrate proteins in vitro, thus enhancing poly-Ubiquitin chain accumulation.  Some DUBs, such as those of the JAMM class, are not inhibited by Ubiquitin-Vinyl Sulfone.

• References:

1. Sharp, P.M. & W.-H. Li. (1987) Trends Ecol. Evol. 2:328.

2. Ciechanover, A. et al. (1980 ) Proc. Natl. Acad. Sci. USA 77:1365.

3. Hershko, A. et al. (1980) Proc. Natl. Acad. Sci. USA 77:1783.

4. Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.

5. Tong, X. et al. (2012) J. Biol. Chem. 287:25280.

6. Wei, W. et al. (2004) Nature 428:194.

7. Wertz, I.E. et al. (2004) Nature 430:694.

8. Borodovsky A., et al. (2001) EMBO J. 20: 5187-5196.

9. Borodovsky A., et al. (2002) Chem. Biol. 9: 1149-1159.

10. Borodovsky A., et al. (2005) Chem. Biochem. 6: 287-291.

11. Hemelaar J., et al. (2004) Mol. Cell. Bio. 24: 84-95.

12. Kessler B.M. (2006) Exp. Rev. Proteomics. 3 :213-221.

13. Misaghi S., et al. (2005) J. Biol. Chem. 280: 1512-1520.

14. Wang G., et al. (2003) Org. Lett. 5: 737-740.

15. Verma R., et al. (2002) Science 298: 611-615.

• Entrez Gene IDs:

  7314 (Human); 298693 (Rat)

• Alternate Names:

  RPS27A; UBA52; UBB ubiquitin B; UBB; UBC; Ubiquitin