Recombinant Human Neurolysin Protein, CF

R&D Systems | Catalog # 3814-ZN

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Neurolysin Protein (3814-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

E. coli

Accession Number

Q9BYT8

Applications

Enzyme Activity
View all Neurolysin Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Neurolysin protein
Ser38-Pro704, with an N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

77 kDa

SDS-PAGE

73 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic peptide substrate, (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Pro-D-Lys(2,4-dinitrophenyl)-OH or Mca-PLGPK(Dnp)-OH.
The specific activity is >70 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

3814-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: Neurolysin

Neurolysin, also known as Oligopeptidase M (mitochondrial peptidase), soluble angiotensin II-binding protein and endopeptidase EC 3.4.24.16 (1, 2), is a homologue of Thimet Ologopeptidase (THOP1), a zinc peptidase of the M3 family that also includes mitochondrial intermediate peptidase. Neurolysin expresses in two forms with the difference of an N-terminal transit peptide that targets the localization to mitochondria (1). The shorter form exists in cytosol. The recombinant human Neurolysin is expressed without the transit peptide and is isolated from the cytosol. Like THOP1, Neurolysin is capable of cleaving a number of vasoactive peptides such as bradykinin and neurotensin (3). All known substrates of THOP1 and Neurolysin contain 17 or fewer amino acids.

References

  1. Serizawa, A. et al. (1995) J. Biol. Chem. 270:2092.
  2. Barett, A.J. and J.M. Chen (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) pp. 356, Elsevier Academic Press, San Diego.
  3. Norman, M.U. et al. (2003) Am. J. Physiol. Heart Circ. Physiol. 284:H1978.

Alternate Names

AGTBP, Neurotensin Endopeptidase, NLN, Oligopeptidase M

Entrez Gene IDs

57486 (Human); 75805 (Mouse); 117041 (Rat)

Gene Symbol

NLN

UniProt

Q9BYT8

Additional Neurolysin Products

Product Documents for Recombinant Human Neurolysin 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 Human Neurolysin Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Human Neurolysin Protein, CF

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Protocols

View specific protocols for Recombinant Human Neurolysin Protein, CF (3814-ZN):

Materials
  • Assay Buffer: 25 mM Tris, 150 mM NaCl, pH 7.5
  • Recombinant Human Neurolysin (rhNeurolysin) (Catalog # 3814-ZN)
  • Substrate: MCA-Pro-Leu-Gly-Pro-D-Lys(DNP)-OH (Bachem, Catalog # M-2270), 2 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 rhNeurolysin to 5 µg/mL in Assay Buffer.
  2. Dilute Substrate to 50 µM in Assay Buffer.
  3. Load into a black well plate 50 µL of 5 µg/mL rhNeurolysin, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 50 µM Substrate.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:
  • rhNeurolysin: 0.25 µg
  • Substrate: 25 µM

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