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APPBP1-UBA3/NEDD8 E1 Protein, Human

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The APPBP1-UBA3/NEDD8 E1 protein is the regulatory subunit in the UBA3-NAE1 E1 enzyme complex and activates NEDD8 by adenylating its C-terminal glycine with ATP. This forms the NEDD8-UBA3 thioester, which is transferred to the catalytic cysteine of UBE2M. APPBP1-UBA3/NEDD8 E1 Protein, Human is a recombinant protein dimer complex containing human-derived APPBP1-UBA3/NEDD8 E1 protein, expressed by E. coli , with tag free. APPBP1-UBA3/NEDD8 E1 Protein, Human, has molecular weight of 60 kDa (APP-BP1) 50 kDa (UBA3).

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  • Species: Human
  • Source: E. coli
  • Storage:
    Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
  • Experimental Validation Results for APPBP1-UBA3/NEDD8 E1 Protein, Human
  • Experimental Validation Results for APPBP1-UBA3/NEDD8 E1 Protein, Human
  • APPBP1-UBA3/NEDD8 E1 Protein, Human Product Image
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  • Biological Activity
  • Technical Parameters
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Biological Activity

Description

The APPBP1-UBA3/NEDD8 E1 protein is the regulatory subunit in the UBA3-NAE1 E1 enzyme complex and activates NEDD8 by adenylating its C-terminal glycine with ATP. This forms the NEDD8-UBA3 thioester, which is transferred to the catalytic cysteine of UBE2M. APPBP1-UBA3/NEDD8 E1 Protein, Human is a recombinant protein dimer complex containing human-derived APPBP1-UBA3/NEDD8 E1 protein, expressed by E. coli , with tag free. APPBP1-UBA3/NEDD8 E1 Protein, Human, has molecular weight of 60 kDa (APP-BP1) 50 kDa (UBA3).

Background

APPBP1-UBA3/NEDD8 E1 protein functions as the regulatory subunit within the dimeric UBA3-NAE1 E1 enzyme complex. In the process of activating NEDD8, E1 initiates by adenylating the C-terminal glycine residue of NEDD8 with ATP. Subsequently, this residue is linked to the catalytic cysteine, resulting in the formation of a NEDD8-UBA3 thioester and free AMP. The final step involves the transfer of NEDD8 to the catalytic cysteine of UBE2M. Crucial for cell cycle progression through the S-M checkpoint, APPBP1-UBA3/NEDD8 E1 plays a vital role in cellular processes. Interestingly, overexpression of NAE1 induces apoptosis by disrupting NEDD8 conjugation. The covalent attachment of NEDD8 to target proteins, known as 'neddylation,' regulates cell growth, viability, and development. Additionally, the binding of TP53BP2 to the regulatory subunit NAE1 acts to decrease neddylation activity, providing further insights into the intricate regulation of this essential cellular process.

Verified Bioactivity

Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein is a member of the NEDD8-activating (E1) enzyme family that is required for the first step of the enzymatic cascade. Subsequently, NEDD8 is coupled to NEDD8-conjugating (E2) enzyme by forming a thioester bond. In this experiment, we used Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein concentration of 80 nM, reaction conditions will need to be optimized for each specific application.

MCE Validation Data

  • Purity - SDS-PAGE

    Purity - SDS-PAGE

    ≥ 95%, as determined by reducing SDS-PAGE.

  • Bioactivity - Enzymatic Activity Assay

    Bioactivity - Enzymatic Activity Assay

    Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein is a member of the NEDD8-activating (E1) enzyme family that is required for the first step of the enzymatic cascade. Subsequently, NEDD8 is coupled to NEDD8-conjugating (E2) enzyme by forming a thioester bond. In this experiment, we used Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein concentration of 80 nM, reaction conditions will need to be optimized for each specific application.

Technical Parameters

  • Species Human
  • Source E. coli
  • Tag Tag Free
  • Accession

    Q13564-1 (M1-L534)&Q8TBC4-1 (M22-S463)

  • Gene ID
    8883
  • Molecular Construction
    • N-term
    • NAE1 (M1-L534)
      Accession # Q13564-1
    • C-term
    • N-term
    • UBA3 (M22-S463)
      Accession # Q8TBC4-1
    • C-term
  • Synonyms

    NAE1; Proto-Oncogene Protein 1; Prev. APPBP1; NEDD8-Activating Enzyme E1 Subunit; APP-BP1; Alternative Protein NAE1; Ula-1; Protooncogene Protein 1; Amyloid Beta Precursor Protein-Binding Protein 1, 59 KDa; A-116A10.1; Amyloid Beta Precursor Protein Bindi

  • Molecular Weight

    60 kDa (APP-BP1) 50 kDa (UBA3)

  • Purity

    ≥ 95%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder.

Formulation

Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O.

Storage & Stability

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Shipping

Room temperature in continental US; may vary elsewhere.

Calculators

Reconstitution Calculator

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

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Dilution Calculator

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
Unit/mg
= 106 ÷
Biological Activity (ED50) Biological Activity (ED50)
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ng/mL
MOQ
Minimum order quantity
100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     ≥ 95%, as determined by reducing SDS-PAGE.

  • 1/16

    MedChemExpress Validation
     Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein is a member of the NEDD8-activating (E1) enzyme family that is required for the first step of the enzymatic cascade. Subsequently, NEDD8 is coupled to NEDD8-conjugating (E2) enzyme by forming a thioester bond. In this experiment, we used Recombinant Human APPBP1/UBA3 Complex (NEDD8 E1) Protein concentration of 80 nM, reaction conditions will need to be optimized for each specific application.

  • APPBP1-UBA3/NEDD8 E1 Protein, Human Product Image
    1/16
  • Experimental Validation Results for APPBP1-UBA3/NEDD8 E1 Protein, Human
  • Experimental Validation Results for APPBP1-UBA3/NEDD8 E1 Protein, Human
  • APPBP1-UBA3/NEDD8 E1 Protein, Human Product Image