Recombinant Human Ubiquitin Vinyl Sulfone Protein, CF

Product Discontinued

Recombinant Human Ubiquitin Vinyl Sulfone Protein, CF Summary

• Details of Functionality: 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 human Ubiquitin protein
  Contains underivatized and C-terminal VS derivatized protein, quantity is by derivatized content
• Accession #: P0CG47
• Protein/Peptide Type: Recombinant Proteins
• Gene: UBB
• Purity: >98%, by HPLC.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 8.6 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• 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.
• Buffer: 2.15 mg/ml (250 μM) in 50 mM MES, pH 6.0
• Purity: >98%, by HPLC.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Ubiquitin Vinyl Sulfone Protein, CF

• RPS27A
• UBA52
• UBB ubiquitin B
• UBB
• UBC
• Ubiquitin

Background

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.

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.

Publications for Ubiquitin (U-202)(11)

Publications using U-202

• Cranford, MT;Dahmen, SN;Cortez, D;Dewar, JM; Leading and lagging strand abasic sites differentially affect vertebrate replisome progression but involve analogous bypass mechanisms Nucleic Acids Res 2025-09-23 [PMID: 41036622] (Bioassay, Xenopus)
• S Zhang, S Zou, D Yin, L Zhao, D Finley, Z Wu, Y Mao USP14-regulated allostery of the human proteasome by time-resolved cryo-EM Nature, 2022-04-27;605(7910):567-574. 2022-04-27 [PMID: 35477760] (Bioassay, Human)
• S Patchett, Z Lv, W Rut, M Békés, M Drag, SK Olsen, TT Huang A molecular sensor determines the ubiquitin substrate specificity of SARS-CoV-2 papain-like protease Cell Reports, 2021-09-08;36(13):109754. 2021-09-08 [PMID: 34547223] (Bioassay, Human)
• A Nishiyama, CB Mulholland, S Bultmann, S Kori, A Endo, Y Saeki, W Qin, C Trummer, Y Chiba, H Yokoyama, S Kumamoto, T Kawakami, H Hojo, G Nagae, H Aburatani, K Tanaka, K Arita, H Leonhardt, M Nakanishi Two distinct modes of DNMT1 recruitment ensure stable maintenance DNA methylation Nat Commun, 2020-03-06;11(1):1222. 2020-03-06 [PMID: 32144273] (Bioassay, Xenopus)
• HB Rycenga, KB Wolfe, ES Yeh, DT Long Uncoupling of p97 ATPase activity has a dominant negative effect on protein extraction Sci Rep, 2019-07-17;9(1):10329. 2019-07-17 [PMID: 31316150] (Bioassay, Xenopus)
• F Liang, AS Miller, S Longerich, C Tang, D Maranon, EA Williamson, R Hromas, C Wiese, GM Kupfer, P Sung DNA requirement in FANCD2 deubiquitination by USP1-UAF1-RAD51AP1 in the Fanconi anemia DNA damage response Nat Commun, 2019-06-28;10(1):2849. 2019-06-28 [PMID: 31253762] (Bioassay, Human)
• L Chen, G Zhu, EM Johns, X Yang TRIM11 activates the proteasome and promotes overall protein degradation by regulating USP14 Nat Commun, 2018-03-26;9(1):1223. 2018-03-26 [PMID: 29581427] (Bioassay, Human)
• M Boselli, BH Lee, J Robert, MA Prado, SW Min, C Cheng, MC Silva, C Seong, S Elsasser, KM Hatle, TC Gahman, SP Gygi, SJ Haggarty, L Gan, RW King, D Finley An inhibitor of the proteasomal deubiquitinating enzyme USP14 induces tau elimination in cultured neurons J. Biol. Chem., 2017-09-26;0(0):. 2017-09-26 [PMID: 28972160] (Enzyme Assay, Human)
• L Yamaguchi, A Nishiyama, T Misaki, Y Johmura, J Ueda, K Arita, K Nagao, C Obuse, M Nakanishi Usp7-dependent histone H3 deubiquitylation regulates maintenance of DNA methylation Sci Rep, 2017-03-03;7(1):55. 2017-03-03 [PMID: 28246399] (Bioassay, Xenopus)
• M Deng, X Yang, B Qin, T Liu, H Zhang, W Guo, SB Lee, JJ Kim, J Yuan, H Pei, L Wang, Z Lou Deubiquitination and Activation of AMPK by USP10 Mol. Cell, 2016-02-11;61(4):614-24. 2016-02-11 [PMID: 26876938] (Western Blot, Human)
• Iguchi , Masahiro, Kujuro , Yuki, Okatsu , Kei, Koyano , Fumika, Kosako , Hidetaka, Kimura , Mayumi, Suzuki , Norihiro, Uchiyama , Shinichi, Tanaka , Keiji, Matsuda , Noriyuki Parkin-catalyzed ubiquitin-ester transfer is triggered by PINK1-dependent phosphorylation. J Biol Chem, 2013-06-10;288(30):22019-32. 2013-06-10 [PMID: 23754282] (Bioassay, Human)

Bioinformatics

• Gene Symbol: UBB
• Uniprot:
  • P0CG47()