Recombinant Human NEDD8 AMC Protein, CF

Product Discontinued

Recombinant Human NEDD8 AMC Protein, CF Summary

• Details of Functionality: Recombinant Human NEDD8 AMC is a fluorogenic substrate for some NEDD8-specific isopeptidases. Release of AMC fluorescence can be monitored with an excitation wavelength of 345 nm and an emission wavelength of 445 nm. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human NEDD8 AMC concentration of 0.1-1 μM.
• Source: E. coli-derived human NEDD8 protein
• Accession #: Q15843
• Protein/Peptide Type: Recombinant Proteins
• Gene: NEDD8
• Purity: >95%, by HPLC.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 8.7 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: Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
• Buffer: X mg/ml (X μM) in 50 mM HEPES pH 7.5, 100 mM NaCl
• Purity: >95%, by HPLC.

Notes

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

Alternate Names for Recombinant Human NEDD8 AMC Protein, CF

• FLJ43224
• MGC104393
• MGC125896
• MGC125897
• NEDD8
• NEDD-8
• Neddylin
• Neural precursor cell expressed developmentally down-regulated protein 8
• neural precursor cell expressed, developmentally down-regulated 8
• Rub1
• Ubiquitin-like protein Nedd8

Background

Neural Precursor Cell Expressed Developmentally Downregulated Gene 8 (NEDD8), also known as Related to Ubiquitin 1 (Rub1), is a 6-8 kDa member of the Ubiquitin family of proteins. Human pro-NEDD8 is 81 amino acids (aa) in length. Pro-NEDD8 contains one Ubiquitin-like domain (aa 1-65) and a 4-5 aa C-terminal propeptide (1,2). Cleavage of the propeptide exposes a C-terminal glycine residue that is used to generate a glycine-lysine intermolecular bond. Mature human NEDD8 shows 100% aa identity to NEDD8 from mouse, rat, and canine. Human NEDD8 is activated by a distinct NEDD8-activating (E1) enzyme, a heterodimeric complex composed of APPBP1 and UBA3 subunits (3). Activated NEDD8 is subsequently transferred to the UBE2M/Ubc12 or UBE2F NEDD8-conjugating (E2) enzymes (4). Through a process termed neddylation, the ROC1/Rbx1 RING Finger E3 ligase transfers NEDD8 to specific substrates (5). For example, transfer of NEDD8 to the Cullin scaffold activates members of the Cullin-RING Ligase (CRL) family of Ubiquitin ligases (E3) (6-8). CRL-induced ubiquitination is known to regulate proteins that are important for cell cycle progression and cell survival (9,10). Physiologically, NEDD8 plays a critical regulatory role in cell proliferation and dysregulation of the NEDD8 pathway has been associated with several cancer pathologies (11,12).

1. Kumar, S. et al. (1993) Biochem. Biophys. Res. Commun. 195:393.
2. Kamitani, T. et al. (1997) J. Biol. Chem. 272:28557.
3. Huang, D.T. & B.A. Schulman (2005) Methods Enzymol. 398:9.
4. Gong, L. & E.T. Yeh (1999) J. Biol. Chem. 274:12036.
5. Ohki, Y. et al. (2009) Biochem. Biophys. Res. Commun. 381:443.
6. Petroski, M.D. & R.T. Deshares (2005) Nat. Rev. Mol. Cell Biol. 6:9.
7. Hjerpe, R. et al. (2012) Biochem. J. 441:927.
8. Calabrese, M.F. et al. (2011) Nat. Struct. Mol. Biol. 18:947.
9. Tateishi, K. et al. (2001) J. Cell. Biol. 155:571.
10. Ohh, M. et al. (2002) EMBO Rep. 3:177.
11. Luo, Z. et al. (2012) Cancer Res. 72:3360.
12. Soucy, T.A. et al. (2010) Genes Cancer 1:708.

Publications for NEDD8 (UL-552)(5)

Publications using UL-552

• M Karpiyevic, S Adjalley, M Mol, DB Ascher, B Mason, GJ van der He, H Laman, H Ovaa, MCS Lee, K Artavanis- Nedd8 hydrolysis by UCH proteases in Plasmodium parasites PLoS Pathog., 2019-10-28;15(10):e1008086. 2019-10-28 [PMID: 31658303] (Bioassay, Human)
• P Paudel, Q Zhang, C Leung, HC Greenberg, Y Guo, YH Chern, A Dong, Y Li, M Vedadi, Z Zhuang, Y Tong Crystal structure and activity-based labeling reveal the mechanisms for linkage-specific substrate recognition by deubiquitinase USP9X Proc. Natl. Acad. Sci. U.S.A., 2019-03-26;0(0):. 2019-03-26 [PMID: 30914461] (Bioassay)
• JF Beckmann, JA Ronau, M Hochstrass A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibility Nat Microbiol, 2017-03-01;2(0):17007. 2017-03-01 [PMID: 28248294] (Bioassay)
• Malene L Urbanus Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen, Legionella pneumophila Mol. Syst. Biol, 2016-12-16;12(12):893. 2016-12-16 [PMID: 27986836] (Bioassay)
• Birol M, Enchev R, Padilla A, Stengel F, Aebersold R, Betzi S, Yang Y, Hoh F, Peter M, Dumas C, Echalier A Structural and biochemical characterization of the Cop9 signalosome CSN5/CSN6 heterodimer. PLoS ONE, 2014-08-21;9(8):e105688. 2014-08-21 [PMID: 25144743] (Bioassay)

Bioinformatics

• Gene Symbol: NEDD8
• Uniprot:
  • Q15843(Human)