Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein, CF

R&D Systems | Catalog # 10237-ZN

His-tag
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Key Product Details

  • R&D Systems CHO-derived Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein (10237-ZN)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

CHO

Accession Number

XP_005557484

Applications

Enzyme Activity
View all Aminopeptidase PILS/ARTS1 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived cynomolgus monkey Aminopeptidase PILS/ARTS1 protein
Ala37-Leu941
with a C-terminal 6-His tag

Purity

>95%, 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.

N-terminal Sequence Analysis

Ala37

Predicted Molecular Mass

104 kDa

SDS-PAGE

95-107 kDa, under reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Leu-AMC.
The specific activity is >800 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein, CF

Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein Enzyme Activity

Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein Enzyme Activity

Recombinant Cynomolgus Monkey Aminopeptidase PILS/ARTS1 His-tag (Catalog # 10237-ZN) is measured by its ability to cleave the fluorogenic peptide substrate, Leu-AMC.
Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein SDS-PAGE

Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein SDS-PAGE

2 μg/lane of Recombinant Cynomolgus Monkey Aminopeptidase PILS/ARTS1 His-tag was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band under reducing conditions at 101 kDa.

Formulation, Preparation, and Storage

10237-ZN
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. 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: Aminopeptidase PILS/ARTS1

ARTS1 encodes a zinc-dependent metalloprotease (1) known as aminopeptidase PILS (Puromycin-Insensitive Leucyl-Specific), ALAP (adipocyte-derived leucine aminopeptidase), TNFR (type 1 tumor necrosis factor receptor) shedding aminopeptidase regulator, or ERAP1 (endoplasmic reticulum aminopeptidase 1). ARTS1 is widely expressed and known to be present in the endoplasmic reticulum. The protein has four domains with a large cavity between domain II and IV that can accommodate large peptide substrates; the second domain contains the active site and the first domain caps the active site and provides binding sites for the amino terminus of the substrate peptides (2). The enzyme has a unique substrate preference amongst the M1-family of aminopeptidases for longer peptides of up to 16 residues (3, 4) that is consistent with its ability to trim antigen precursors for MHC class 1 presentation (5). Variants and polymorphisms identified in ARTS-1 have linked it to autoimmune disease (6), cancer (7), and hypertension (8). ARTS-1 has been shown to be secreted (9, 10) and cause activation of inflammasome and cathepsin B pathways (11) and consequently to have a direct role in innate immunity and contribute to autoinflammatory and autoimmune diseases (12). ARTS-1 is consequently a potential therapeutic target (12-14).

References

  1. Schomburg, L. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 311, Academic Press, San Diego.
  2. Nguyen, T. T. et al. (2011) Nat. Struct. Mol. Biol. 18:604.
  3. York, I. A. et al. (2002) Nat. Immunol. 3:1177.
  4. Chang, S. C. et al. (2005) Proc. Nat. Acad. Sci. U.S.A. 102:17107.
  5. Serwold, T. et al. (2002) Nature 419:480.
  6. Fung, E. Y. et al. (2009) Genes Immun. 10:188.
  7. Mehta, A. M. et al. (2009) Genes Chromosomes Cancer 48:410.
  8. Yamamoto, N. et al. (2002) Hum. Mutat. 19:251.
  9. Cui, X. et al. (2002) J. Clin. Invest. 110:515.
  10. Goto, Y. et al. (2011) J. Biol. Chem. 286:21906.
  11. Aldhamen, Y. A. et al. (2015) J. Innate Immun. 7:275.
  12. Pepelyayeva, Y. and A. Amalfitano. (2019) Hum. Immunol. 80:302.
  13. Koumantou, D. et al. (2019) Cancer Immunol. Immunother. 68:1245.
  14. Georgiadis, D. et al. (2018) Curr. Med. Chem. [Epub ahead of print]

Long Name

Aminopeptidase Puromycin-insensitive Leucyl-specific

Alternate Names

A-LAP, ARTS1, ERAAP1, ERAP1, PILS-AP

Entrez Gene IDs

51752 (Human); 80898 (Mouse); 102146570 (Cynomolgus Monkey)

Gene Symbol

ERAP1

UniProt

XP_005557484

Additional Aminopeptidase PILS/ARTS1 Products

Product Documents for Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

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Protocols

View specific protocols for Recombinant Cynomolgus Aminopeptidase PILS/ARTS1 Protein, CF (10237-ZN):

Materials
  • Assay Buffer: 25 mM Tris, pH 8.0
  • Recombinant Cynomolgus Monkey Aminopeptidase PILS/ARTS1 His-tag (cynoARTS1) (Catalog # 10237-ZN)
  • Substrate: Leu-AMC (Bachem, Catalog # I-1240), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rcynoARTS1 to 1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 600 µM in Assay Buffer.
  3. Load in plate 50 µL of 1 µg/mL rcynoARTS1, and start the reaction by adding 50 µL of 600 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 600 µM Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode of 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 AMC (Sigma, Catalog # A9891)

Per Well:
  • rcynoARTS1: 0.05 µg
  • Substrate: 300 µM

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