详细说明
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Activity
Recombinant Human NEDD8 can be conjugated to substrate proteins via the subsequent actions of a NEDD8-activating (E1) enzyme, a NEDD8-conjugating (E2) enzyme, and a NEDD8 ligase (E3). Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human NEDD8 concentration of 10-50 μM.
Source
E. coli-derived
Accession #
Predicted Molecular Mass
8.6 kDa
UL-812 |
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Formulation X mg/ml (X μM) in 50 mM Hepes, pH 8, 150mM NaCl, and 1mM DTT | ||
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.
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Background: NEDD8
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).
References:
12. Soucy, T.A.
et al. (2010) Genes Cancer
1:708.
11. Luo, Z.
et al. (2012) Cancer Res.
72:3360.
10. Ohh, M.
et al. (2002) EMBO Rep.
3:177.
9. Tateishi, K.
et al. (2001) J. Cell. Biol.
155:571.
8. Calabrese, M.F.
et al. (2011) Nat. Struct. Mol. Biol.
18:947.
7. Hjerpe, R.
et al. (2012) Biochem. J.
441:927.
6. Petroski, M.D. & R.T. Deshares (2005) Nat. Rev. Mol. Cell Biol.
6:9.
5. Ohki, Y.
et al. (2009) Biochem. Biophys. Res. Commun.
381:443.
4. Gong, L. & E.T. Yeh (1999) J. Biol. Chem.
274:12036.
3. Huang, D.T. & B.A. Schulman (2005) Methods Enzymol.
398:9.
1. Kumar, S.
et al.(1993) Biochem. Biophys. Res. Commun.
195:393. 2. Kamitani, T.
et al. (1997) J. Biol. Chem.
272:28557.
Long Name:
Neural Precursor Cell Expressed Developmentally Down-Regulated Protein 8
Entrez Gene IDs:
4738 (Human); 18002 (Mouse); 25490 (Rat)
Alternate Names:
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