Recombinant Human Aminopeptidase PILS/ARTS1 Protein, CF

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

Recombinant Human Aminopeptidase PILS/ARTS1 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate, Leu-AMC. The specific activity is >125 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Aminopeptidase PILS/ARTS1 protein
  Ala37-Met941, with a C-terminal 10-His tag
• Accession #: EAW96077
• N-terminal Sequence: Ala37
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ERAP1
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 105 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: 108 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 Tris and NaCl.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 25 mM Tris, pH 8.0
  • Recombinant Human Aminopeptidase PILS/ARTS1 (rhARTS1) (Catalog # 2334-ZN)
  • Substrate: Leu-AMC (Bachem, Catalog # I-1240)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhARTS1 to 2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 600 µM in Assay Buffer.
  3. Load 50 µL of 2 µg/mL rhARTS1 into the plate, and start the reaction by adding 50 µL of 600 µM Substrate. Include a Substrate Blank containing Assay Buffer and 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:
  • rhARTS1: 0.1 µg
  • Substrate: 300 µM

Notes

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

Alternate Names for Recombinant Human Aminopeptidase PILS/ARTS1 Protein, CF

• Adipocyte-derived leucine aminopeptidase
• A-LAP
• A-LAPALAP
• Aminopeptidase PILS
• ARTS1
• ARTS1aminopeptidase regulator of TNFR1 shedding
• ARTS-1PILSAP
• EC 3.4.11
• EC 3.4.11.-
• EC 3.4.11.1
• endoplasmic reticulum aminopeptidase 1
• endoplasmic reticulum aminopeptidase associated with antigen processing
• ERAAP1
• ERAP1
• KIAA0525APPILS
• PILS-AP
• PILS-APERAAP
• Puromycin-insensitive leucyl-specific aminopeptidase
• Type 1 tumor necrosis factor receptor shedding aminopeptidase regulator

Background

The name of Aminopetidase PILS (Puromycin-Insensitive Leucyl-Specific) describes the two basic properties of this zinc metalloprotease in vitro (1). Also known as ALAP (Adipocyte-derived Leucin AminoPeptidase), type 1 tumor necrosis factor receptor (TNFR) shedding aminopeptidase regulator and ER aminopeptidase ERAP1 or ERAAP, it is encoded by the ARTS1 gene (2-4). Aminopeptidase PILS has been identified to regulate antigen presentation, promote TNFR1 ectodomain shedding and associate with hypertension (2-5).

1. Schomburg, L. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 311, Academic Press, San Diego.
2. Cui, X. et al. (2002) J. Clin. Invest. 110:515.
3. York, I.A. et al. (2002) Nat. Immunol. 3:1177.
4. Serwold, T. et al. (2002) Nature 419:480.
5. Yamamoto, N. et al. (2002) Hum. Mutat. 19:251.

Publications for Aminopeptidase PILS/ARTS1 (2334-ZN)(4)

Publications using 2334-ZN

• M Laura, G Ronan, LB Vy, G Valentin, CA Omar, C Virgyl, C Piveteau, R Melissa, F Charlotte, W Sandrine, C Julie, DR Julie, B Damien, L Florence, VE Peter, D Benoit, DP Rebecca Modulators of hERAP2 discovered by high-throughput screening European Journal of Medicinal Chemistry, 2020-12-11;211(0):113053. 2020-12-11 [PMID: 33359953] (Bioassay)
• K Textoris-T, C Cammann, P Henklein, E Topfstedt, F Ebstein, S Henze, J Liepe, F Zhao, D Schadendor, B Dahlmann, W Uckert, A Paschen, M Mishto, U Seifert ER-aminopeptidase 1 determines the processing and presentation of an immunotherapy-relevant melanoma epitope Eur. J. Immunol., 2019-11-27;0(0):. 2019-11-27 [PMID: 31729751] (Enzyme Assay, Human)
• Kourjian , Georgio, Xu , Yang, Mondesire-Crump , Ijah, Shimada , Mariko, Gourdain , Pauline, Le Gall , Sylvie Sequence-specific alterations of epitope production by HIV protease inhibitors. J Immunol, 2014-03-10;192(8):3496-506. 2014-03-10 [PMID: 24616479] (Enzyme Assay, Human)
• Krige D, Needham LA, Bawden LJ, Flores N, Farmer H, Miles LE, Stone E, Callaghan J, Chandler S, Clark VL, Kirwin-Jones P, Legris V, Owen J, Patel T, Wood S, Box G, Laber D, Odedra R, Wright A, Wood LM, Eccles SA, Bone EA, Ayscough A, Drummond AH CHR-2797: an antiproliferative aminopeptidase inhibitor that leads to amino acid deprivation in human leukemic cells. Cancer Res., 2008-08-15;68(16):6669-79. 2008-08-15 [PMID: 18701491] (Bioassay, Human)

Bioinformatics

• Gene Symbol: ERAP1
• Uniprot:
  • EAW96077()