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

R&D Systems, part of Bio-Techne | Catalog # E-313B

His-tag
R&D Systems, part of Bio-Techne
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E-313B-025

Key Product Details

Source

Sf 21 (baculovirus)

Structure / Form

Complex

Conjugate

Unconjugated

Applications

Enzyme Activity

Product Specifications

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)

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Predicted Molecular Mass

62 kDa (APPBP1) & 52 kDa (UBA3)

SDS-PAGE

57-63 kDa (APPBP1) & 46-54 kDa (UBA3), under reducing conditions

Activity

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.

Scientific Data Images for Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein, CF

Recombinant Human APPBP1/UBA3 Complex (NEDD8 Activating Enzyme) His-tag Protein SDS-PAGE.

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.

Formulation, Preparation and Storage

E-313B
Formulation Supplied as a 0.2 μm filtered solution in HEPES, NaCl, and 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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: APPBP1/UBA3 Complex

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).





References

  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.

Long Name

Amyloid beta Precursor Protein-binding Protein 1/Ubiquitin Like Modifier Activating Enzyme 3

Additional APPBP1/UBA3 Complex Products

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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein, CF

For research use only

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