Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein, CF

2 μg/lane of Recombinant Human APPBP1/UBA3 Complex (NEDD8 Activating Enzyme) His-tag Protein (Catalog # E-313B) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 57-63 kDa (APPBP1) & 46-54 kDa (UBA3), under reducing conditions.

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein, CF Summary

• Additional Information: His-tag
• Details of Functionality: Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein concentration of 50-200 nM.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived human APPBP1/UBA3 Complex protein
  Ala2-Leu534 with an N-terminal Met and 6-His tag (APPBP1), Met1-Ser463 (UBA3)
• Accession #: Q13564.1(APPBP1) Q8TBC4.2 (UBA3)
• Structure / Form: Complex
• Protein/Peptide Type: Recombinant Enzymes
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 62 kDa (APPBP1) & 52 kDa (UBA3).
  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.
• SDS-PAGE: 57-63 kDa (APPBP1) & 46-54 kDa (UBA3), under reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in HEPES, NaCl, and DTT.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining

Notes

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

Alternate Names for Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein, CF

• A-116A10.1
• amyloid beta precursor protein binding protein 1
• amyloid beta precursor protein binding protein 1, 59kDa
• Amyloid beta precursor protein-binding protein 1, 59 kDa
• amyloid beta precursor protein-binding protein 1, 59kD
• Amyloid protein-binding protein 1
• APPBP1/UBA3 Complex
• APPBP1APP-BP1
• NEDD8 activating enzyme E1 subunit 1
• NEDD8-activating enzyme E1 regulatory subunit
• NEDD8-activating enzyme E1 subunit
• protooncogene protein 1
• Proto-oncogene protein 1
• ula-1

Background

Recombinant Human APPBP1/UBA3 Complex (NEDD8 Activating Enzyme) is a heterodimeric enzyme with a predicted molecular weight of 112 kDa and member of the NEDD8-activating (E1) enzyme family. It is responsible for the first step in the enzymatic cascade that also utilizes a NEDD8-conjugating (E2) enzyme and a NEDD8 ligase (E3) in order to conjugate NEDD8 to protein substrates. The heterodimer is composed of a regulatory subunit, Amyloid beta Precursor Protein Binding Protein 1 (APPBP1), and a catalytic subunit, Ubiquitin-like Modifier Activating Enzyme 3 (UBA3). Human APPBP1 is a 534 amino acid (aa) protein with a predicted molecular weight of 60 kDa that is expressed ubiquitously in fetal tissues and in the adult brain (1). APPBP1 is required for UBA3 neddylation activity, regulates enzyme specificity, and is expressed as two isoforms, the full length protein and a second isoform with an alternate N-terminal, aa1-17, sequence (2). APPBP1 has been shown to drive cell cycle progression, and its expression is increased in the hippocampus of Alzheimer's disease brains (3, 4). Human UBA3 is a 463 aa protein with a predicted molecular weight of 52 kDa. It is ubiquitously expressed and shares high aa sequence identity with the C-terminal domain of human UBE1 (5). UBA3 contains an ATP-binding domain and an active site cysteine residue, Cys237 in humans, which are both common to E1 enzymes. Like APPBP1, two isoforms of UBA3 have been identified in humans, the full length protein and a truncated isoform, which lacks aa 8-21. UBA3 is required for cell cycle progression and has been shown to downregulate steroid receptor activation (4, 6). Neddylation and its associated enzymes have been implicated in the progression of Alzheimer's disease, via neddylation of APP, and cancer via post-translational modification of oncogenes (7, 8).

1. Chow, N. et al. (1996) J. Biol. Chem. 271:11339.
2. Walden, H. et al. (2003) Mol. Cell 12:1427.
3. Chen, Y. et al. (2003) J. Cell Biol. 163:27.
4. Chen, Y. et al. (2000) J. Biol. Chem. 275:8929.
5. Gong, L. & E.T. Yeh (1999) J. Biol. Chem. 274:12036.
6. Fan, M. et al. (2002) Mol. Endocrinol. 16:315.
7. Chen, Y. et al. (2012) J. Cell Mol. Med. 16:2583.
8. Soucy, T.A. et al. (2010) Genes Cancer 1:708.

Publications for NEDD8 Activating Enzyme (APPBP1/UBA3) (E-313B)(1)

Publication using E-313B

• Iwatani, Y;Matsuoka, K;Ode, H;Kubota, M;Nakata, Y;Setoyama, Y;Kojima, K;Imahashi, M;Yokomaku, Y; The unique structure of the highly conserved PPLP region in HIV-1 Vif is critical for the formation of APOBEC3 recognition interfaces mBio 2025-01-21 [PMID: 39835817] (Bioassay, N/A)

Bioinformatics

• Uniprot:
  • Q13564.1(APPBP1) Q8TBC4.2 (UBA3)()