Recombinant Human Aminopeptidase A/ENPEP Protein, CF

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

Recombinant Human Aminopeptidase A/ENPEP Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate, Glu-7-amido-4-methylcoumarin (Glu-AMC). The specific activity is > 2,000 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Aminopeptidase A/ENPEP protein
  Arg41-Gly957, with an N-terminal 6-His tag
• Accession #: AAA35522
• N-terminal Sequence: His
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ENPEP
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 106 kDa.
  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: 145 kDa, 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 MES and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Buffer: 25 mM Tris, 50 mM CaCl₂, 0.2 M NaCl, pH 8.0
  • Recombinant Human Aminopeptidase A/ENPEP (rhENPEP) (Catalog # 2499-ZN)
  • Substrate: Glu-AMC (Bachem, Catalog # I-1180), 10 mM stock in 30 mM HCl
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhENPEP to 0.4 ng/µL in Assay Buffer.
  2. Dilute Substrate Glu-AMC to 200 µM in Assay Buffer.
  3. Load into plate 50 µL of 0.4 ng/µL rhENPEP, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)) / (amount of enzyme (µg))

  *Adjusted for Substrate Blank
  **Derived using calibration standard 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

  Per Well:
  • rhENPEP: 0.020 μg
  • Substrate: 100 µM

Notes

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

Alternate Names for Recombinant Human Aminopeptidase A/ENPEP Protein, CF

• Aminopeptidase A
• APA
• AP-A
• CD249 antigen
• CD249
• Differentiation antigen gp160
• EAP
• EC 3.4.11
• EC 3.4.11.7
• ENPEP
• glutamyl aminopeptidase (aminopeptidase A)
• glutamyl aminopeptidase
• gp160

Background

The human ENPEP gene encodes aminopeptidase A (APA), which is also known as glutamyl aminopeptidase (EAP), aspartate aminopeptidase, angiotensinase A, Ca²⁺-activated glutamate aminopeptidase, membrane aminopeptidase II and the BP-1/6C3 antigen (1). The various names reflect its substrate specificity (with a preference for Glu, Asp and angiotensin II as one of the physiological substrates), Ca²⁺-dependence, cellular location (type II membrane protein) and immunological reactivity to the antibody BP-1/6C3. The deduced amino acid sequence of human ENPEP consists of a short cytoplasmic tail (residues 1 to 17), a transmembrane region (residues 18 to 40), and a long ectodomain (residues 41 to 957) (2, 3). In addition to the N-terminal zinc metalloprotease domain, the ectodomain also contains the C-terminal region, which functions as an intramolecular chaperone required for the correct folding, cell surface expression and activity (4). The purified recombinant human ENPEP consists of the entire ectodomain and is active in the assay as described in Activity Assay Protocol.

1. Wang, J. and M.D. Cooper (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) p. 299, Academic Press, San Diego.
2. Nanus, D.M. et al. (1993) Proc. Natl. Acad. Sci. USA 90:7069.
3. Li, L. et al. (1993) Genomics 17:657.
4. Rozenfeld, R. et al. (2004) J. Biol. Chem. 279:43285.

Publications for Aminopeptidase A/ENPEP (2499-ZN)(2)

Publications using 2499-ZN

• Forster, P;Wysocki, J;Abedini, Y;Müller, T;Ye, M;Ferrario, CM;Batlle, D; Aminopeptidase A Effect on Angiotensin Peptides and Their Blood Pressure Action International journal of molecular sciences 2025-07-21 [PMID: 40725237] (In vitro assay, N/A)
• Sevalle J, Amoyel A, Robert P, Fournie-Zaluski MC, Roques B, Checler F Aminopeptidase A contributes to the N-terminal truncation of amyloid beta-peptide. J. Neurochem., 2009-02-23;109(1):248-56. 2009-02-23 [PMID: 19187443] (Enzyme Assay, Human)

Bioinformatics

• Gene Symbol: ENPEP
• Uniprot:
  • AAA35522()