Recombinant Human NPEPPS Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Human NPEPPS Protein, CF Summary

Details of Functionality
Measured by its ability to cleave the fluorogenic peptide substrate, Leu-AMC. The specific activity is >2,500 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Puromycin-sensitive aminopeptidase/NPEPPS protein
Pro46-Val919, with an N-terminal Met and a C-terminal 10-His tag
Accession #
N-terminal Sequence
Pro46
Protein/Peptide Type
Recombinant Enzymes
Gene
NPEPPS
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
100 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
90-100 kDa, reducing conditions
Publications
Read Publication using
6410-ZN in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Assay Procedure
  • Assay Buffer: 25 mM HEPES, 1 mM DTT, pH 7.0
  • Recombinant Human Puromycin‑sensitive aminopeptidase/NPEPPS (rhNPEPPS) (Catalog # 6410-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 rhNPEPPS to 0.2 ng/μL in Assay Buffer.
  2. Dilute Substrate to 20 μM in Assay Buffer.
  3. Load 50 μL of 0.2 ng/μL rhNPEPPS into the microplate and start the reaction by adding 50 μL of Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, for 5 minutes in kinetic mode.
  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:
  • rhNPEPPS: 0.010 µg
  • Substrate: 10 µM

Notes

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.


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

Alternate Names for Recombinant Human NPEPPS Protein, CF

  • AAP-S
  • aminopeptidase puromycin sensitive
  • Cytosol alanyl aminopeptidase
  • EC 3.4.11.2
  • EC 3.4.11.3
  • metalloproteinase MP100
  • MP100
  • MP100EC 3.4.11
  • NPEPPS
  • PSAEC 3.4.11.14
  • puromycin-sensitive aminopeptidase

Background

NPEPPS, also known as Puromycin-sensitive aminopeptidase and cytosol alanyl aminopeptidase, is a zinc metallopeptidase which hydrolyzes N‑terminal amino acids from its substrates. NPEPPS is expressed in most tissues as a cytoplasmic protein, but a membrane-associated form has been identified in the brain (1, 2). The enzyme has broad substrate specificity but prefers basic and hydrophobic residues in the substrate P1 and P'1 sites (3). NPEPPS is known to hydrolyze dynorphins and enkephalins, and may contribute to the degradation of these peptides in the brain (4). NPEPPS plays a major role in the degradation of toxic polyglutaminyl peptides that are released by proteasomes (5). NPEPPS is also known to degrade the tau protein, which accumulates and polymerizes in some neurodegenerative diseases (6).
  1. Dyer, S.H. et al. (1990) J. Neurochem. 54:547.
  2. McLellan, S. et al. (1988) J. Neurochem. 51:1552.
  3. Johnson, G.D. and L.B. Hersh (1990) Arch. Biochem. Biophys. 276:305.
  4. Safavi, A. and L.B. Hersh (1995) J. Neurochem. 65:389.
  5. Bhutani, N. et al. (2007) EMBO J. 26:1385.
  6. Sengupta, S. et al. (2006) Biochemistry 45:15111.

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Publications for Puromycin-sensitive aminopeptidase/NPEPPS (6410-ZN)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: Bioassay.


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Bioassay
(1)
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(1)
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Bioinformatics

Gene Symbol NPEPPS
Entrez
Uniprot