Recombinant Human NEDD8 Protein

Name: Recombinant Human NEDD8 Protein
Brand: Beta LifeScience
SKU: BL-2759NP
Availability: InStock

BL-2759NP: Greater than 95% as determined by reducing SDS-PAGE. (QC verified)

Collections: All products, High-quality recombinant proteins, Other recombinant proteins

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Product Overview

Description	Recombinant Human Neural precursor cell expressed developmentally down-regulated protein 8 is produced by our E.coli expression system and the target gene encoding Met1-Gly76 is expressed.
Accession	Q15843
Synonym	Neural precursor cell expressed developmentally down-regulated protein 8; NEDD8; Neddylin; Ubiquitin-like protein Nedd8
Gene Background	Human NEDD8 shares 60% amino acid sequence identity to ubiquitin. The only known substrates of NEDD8 modification are the cullin subunits of SCF ubiquitin E3 ligases. The NEDDylation of cullins is critical for the recruitment of E2 to the ligase complex, thus facilitating ubiquitin conjugation. NEDD8 modification has therefore been implicated in cell cycle progression and cytoskeletal regulation.
Molecular Mass	8.6 KDa
Apmol Mass	9 KDa, reducing conditions
Formulation	Lyophilized from a 0.2 μm filtered solution of 20mM Tris-HCl, 250mM NaCl, pH 8.0.
Endotoxin	Less than 0.1 ng/µg (1 EU/µg) as determined by LAL test.
Purity	Greater than 95% as determined by reducing SDS-PAGE. (QC verified)
Biological Activity	Not tested
Reconstitution	Always centrifuge tubes before opening. Do not mix by vortex or pipetting. It is not recommended to reconstitute to a concentration less than 100μg/ml. Dissolve the lyophilized protein in distilled water. Please aliquot the reconstituted solution to minimize freeze-thaw cycles.
Storage	Lyophilized protein should be stored at ≤ -20°C, stable for one year after receipt. Reconstituted protein solution can be stored at 2-8°C for 2-7 days. Aliquots of reconstituted samples are stable at ≤ -20°C for 3 months.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature listed below.
Usage	For Research Use Only

Target Details

Target Function	Ubiquitin-like protein which plays an important role in cell cycle control and embryogenesis via its conjugation to a limited number of cellular proteins, such as cullins or p53/TP53. Attachment of NEDD8 to cullins is critical for the recruitment of E2 to the cullin-RING-based E3 ubiquitin-protein ligase complex, thus facilitating polyubiquitination and proteasomal degradation of cyclins and other regulatory proteins. Attachment of NEDD8 to p53/TP53 inhibits p53/TP53 transcriptional activity. Covalent attachment to its substrates requires prior activation by the E1 complex UBE1C-APPBP1 and linkage to the E2 enzyme UBE2M.
Subcellular Location	Nucleus.
Protein Families	Ubiquitin family
Database References	HGNC: 7732 OMIM: 603171 KEGG: hsa:4738 STRING: 9606.ENSP00000250495 UniGene: PMID: 28169289 NEDD8-mediated neddylation promotes stress granule assembly. PMID: 27381497 NEDD8 seems to inhibit the Src-mediated phosphorylation of caveolin-1 by modifying the structure of caveolin-1 protein, which blocks the migration of cancer cells. Although the neddylation process is currently regarded as an emerging target for cancer therapy, our results suggest the possibility that the inhibition of neddylation could facilitate cancer invasion or metastasis at least in some types of cancers. PMID: 29301501 The cellular effects of NEDD8 inhibition can be manipulated based on the p53 status. PMID: 27901050 Here we report that high expression of neddylation components, NEDD8 and NAE1, are associated with poor survival of Pancreatic ductal adenocarcinoma patients. Blockage of neddylation by MLN4924 significantly sensitizes pancreatic cancer cells to gemcitabine, reduced clonogenic survival, decreased invasion capacity, increased apoptosis and senescence PMID: 28535453 Downregulation of NEDD8 enhanced the susceptibility of nasopharyngeal carcinoma cells to cisplatin and radiation. PMID: 28569775 Furthermore, the authors characterized SENP8/DEN1 as the protease that counteracts Ubc12 auto-neddylation, and observed aberrant neddylation of Ubc12 and other NEDD8 conjugation pathway components in SENP8-deficient cells. PMID: 28475037 MKK7 undergoes neddylation in human breast cancer cells PMID: 26364603 complete blocking of CRLs at a higher inhibitor dose-induced cytotoxicity that was amplified by knockdown of CRL regulator Cand1. PMID: 27906189 In conclusion, the authors revealed that E3 ligase HDM2 promotes NEDD8 NEDDylation of HBx to enhance HBx stability and chromatin localization, which in turn favors HBx-dependent transcriptional regulation, cell proliferation, and hepatitis B virus-driven tumor growth. PMID: 28592528 CUL5 neddylation may allosterically tune polyubiquitin chain length and topology. PMID: 28082425 This study identifies MyD88 as a novel substrate of NEDD8, and demonstrate that MyD88 NEDDylation antagonizes its ubiquitination. PMID: 27864145 This study provides new knowledge of the CFTR biosynthetic pathway. It suggests that SYVN1 and FBXO2 represent two distinct multiprotein complexes that may degrade DeltaF508-CFTR in airway epithelia and identifies a new role for NEDD8 in regulating DeltaF508-CFTR ubiquitination. PMID: 27756846 Neddylation of Smurf1 activates its ubiquitin ligase activity and Smurf2 exerts Nedd8 ligase activity. This study provided new clues of Smurf2 activation regulation. PMID: 27086113 Nedd8(Q40E) cannot induce the same structural effect on Cul1-Rbx1 as wild-type Nedd8. PMID: 26632597 findings add novel evidence showing, for what we believe the first time, that NEDD8-mediated neddylation is required for normal human endometrial functions PMID: 26003431 In endothelial dysfunction, HDAC2 levels were reciprocally regulated by ectopic expression of NEDD8 and the de-NEDDylating enzyme SENP8. PMID: 25655932 we report that MLN4924 (NEDD8 enzyme inhibitor ) specifically inhibited protein neddylation, inactivated cullin-RING E3 ligase (CRL), the best-known neddylation substrate, and induced the accumulation CRL substrates in lymphoma cells. PMID: 25782162 E1 (NAE1 and UBA3) and E2 (UBC12) enzymes, as well as global NEDD8 conjugation, were upregulated in over 2/3 of human intrahepatic cholangiocarcinoma PMID: 25229838 This work sheds new light on the roles of NEDD8 lysines on neddylation cascades and provides a dominant negative mutant for the study of neddylation and its biological functions. PMID: 25918018 a role of Nedd8 in regulating caspase-1 activation following inflammasome activation PMID: 25452302 NEDD8 negatively regulates the DNA damage repair process through suppression of the ubiquitylation of H2A and gammaH2AX, which further blocks the recruitment of the damage response protein BRCA1. PMID: 24634510 SCCRO3 functions as a tumor suppressor by antagonizing the neddylation activity of SCCRO PMID: 25349211 Study reports the structure of a trapped RING E3-E2 approximately UBL-target intermediate representing RBX1-UBC12 approximately NEDD8-CUL1-DCN1, which reveals the mechanism of NEDD8 ligation and how a particular UBL and acceptor lysine are matched by a multifunctional RING E3. PMID: 24949976 Protein neddylation with NEDD8 protein plays an important role in HIV-1 and HIV-2 infection. PMID: 24245672 More importantly, RPS14 was specifically modified with NEDD8 and hCINAP inhibited RPS14 NEDDylation by recruiting NEDD8-specific protease 1 PMID: 23246961 Our results suggest that NEDD8 may play an important role in the control of proliferation and differentiation of human placenta throughout pregnancy. PMID: 23812220 In coordination with the P97-UFD1-NPL4 complex (P97(UFD1/NPL4)), NUB1L promotes transfer of NEDD8 to proteasome for degradation. PMID: 24019527 these results indicate that NAE1/APP-BP1 and NEDDylation are invovled in modulating p53 activity and regulating its role in the response of cells to ionizing radiation PMID: 22895816 Data indicate that overexpression of SENP8, a NEDD8-specific cysteine protease, resulted in deNEDDylation of E2F1 and promoted its transactivation activity at the p73 gene. PMID: 23001041 These studies demonstrate that disrupting host NEDD8 cascades presents a novel antiretroviral therapeutic approach enhancing the ability of the immune system to combat HIV. PMID: 23300442 these findings suggest that once Rub1/Nedd8 is channeled into ubiquitin pathways, it is recognized essentially like ubiquitin. PMID: 23105008 NEDD8 modifies transcription factor E2F-1. PMID: 22836579 histone H4 was polyneddylated in response to DNA damage, and NEDD8 was conjugated to the N-terminal lysine residues of H4 PMID: 23394999 a new regulatory mechanism of RCAN1 function PMID: 23118980 c-Cbl conjugates neural precursor cell-expressed, developmentally downregulated 8 (NEDD8), a ubiquitin-like protein, to TbetaRII at Lys556 and Lys567. PMID: 23290524 Neddylation is achieved via a multienzyme process wherein SENP8 enables cleavage of the Nedd8 precursor and promotes Nedd8 conjugation to the Cullin-RING ligases. PMID: 23209320 present a view of how a bacterial deamidase effector, cycle-inhibiting factor homolog in Burkholderia pseudomallei, recognizes its host targets, ubiquitin (Ub) and Ub-like NEDD8, and catalyzes site-specific deamidation PMID: 23175788 Deconjugation of Nedd8 from Cul1 is directly regulated by Skp1-F-box and substrate, and the COP9 signalosome inhibits deneddylated SCF by a noncatalytic mechanism. PMID: 22767593 In the present study, the proportions of intranuclear inclusion positive for NEDD8, NUB1 and SUMO-1 were significantly lower in glial cells than in neurons. PMID: 22612509 We report the crystal structures of two Cif/NEDD8 complexes, revealing a conserved molecular interface that defines enzyme/substrate recognition. PMID: 22691497 Decrease in free ubiquitin levels under stress allows NEDD8 to be conjugated through Ube1. PMID: 22370482 The studies provide insights on how nucleolar stress through L11 and NEDD8 can activate the transcriptional activity of p53. PMID: 22081073 High NEDD8 is associated with melanoma. PMID: 22502714 XIAP does not function as a NEDD8-E3 ligase for caspase-7 in vivo PMID: 22584050 These results uncover an unexpected and conserved role for NEDD8 in linking cullin-RING ubiquitin ligases ubiquitin ligase function to the p97 pathway. PMID: 22466964 Mdm2/NEDD8/HuR regulatory framework is essential for the malignant transformation of tumor cells. PMID: 22095636 E1-E2 interactions in ubiquitin and Nedd8 ligation pathways PMID: 22069333 In cells, NEDD8 overexpression leads to this type of NEDDylation by increasing the concentration of NEDD8, whereas proteasome inhibition has the same effect by depleting free ubiquitin. PMID: 22004789 findings suggest that TRIM40 inhibits NF-kappaB activity via neddylation of inhibitor of nuclear factor kappaB kinase subunit gamma and that TRIM40 prevents inflammation-associated carcinogenesis in the gastrointestinal tract PMID: 21474709

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

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