Neprilysin/CD10, also known as NEP and neutral endopeptidase 24.11, is a zinc metallopeptidase expressed at the cell surface of a variety of cells. The enzyme functions both as an endopeptidase with a thermolysin-like specificity and as a dipeptidylcarboxypeptidase. NEP has been shown to be involved in the degradation of enkephalins in the mammalian brain and the inactivation of circulating atrial natriuretic peptide (1, 2). NEP has also been identified as the common acute lymphocytic leukemia antigen (CALLA), and is expressed on the surface of lymphocytes in some disease states (3, 4). These and other observations have resulted in considerable interest in NEP as a target for analgesics and antihypertensive drugs. NEP is also a major degrading enzyme of amyloid beta peptide (A beta ) in the brain, indicating that down-regulation of NEP activity, which could be caused by aging, can contribute to the development of Alzheimer’s disease by promoting A beta accumulation (5).
Recombinant Human Neprilysin (CHO-expressed) Protein, CF
R&D Systems | Catalog # 1182-ZNC
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Key Product Details
- R&D Systems CHO-derived Recombinant Human Neprilysin (CHO-expressed) Protein (1182-ZNC)
- 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
Applications
Enzyme Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human Neprilysin/CD10 protein
Tyr52-Trp750, with an N-terminal 6-His tag
Tyr52-Trp750, with an N-terminal 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
His
Predicted Molecular Mass
81 kDa
SDS-PAGE
102 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005).
The specific activity is >1,500 pmol/min/µg, as measured under the described conditions.
The specific activity is >1,500 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1182-ZNC
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and ZnCl2. |
| 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.
|
Background: Neprilysin/CD10
References
- Malfroy, B. et al. (1978) Nature 276:523.
- Kenny, A.J. and Stephenson, S.L. (1988) FEBS Lett. 232:1.
- LeTarte, M. et al. (1988) J. Exp. Med. 168:1247.
- Shipp, M.A. et al. (1988) Proc. Natl. Acad. Sci. USA 85:4819.
- Itwata, N. et al. (2001) Science 292:1550.
Alternate Names
CALLA, CD10, Enkephalinase, Leu-19, MME, Neutral Endopeptidase 24.11, NKH1
Gene Symbol
MME
UniProt
Additional Neprilysin/CD10 Products
Product Documents for Recombinant Human Neprilysin (CHO-expressed) Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Neprilysin (CHO-expressed) Protein, CF
For research use only
Citations for Recombinant Human Neprilysin (CHO-expressed) Protein, CF
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Protocols
View specific protocols for Recombinant Human Neprilysin (CHO-expressed) Protein, CF (1182-ZNC):
Materials
- Assay Buffer: 50 mM Tris, 0.05% Brij-35, pH 9.0
- Recombinant Human Neprilysin (rhNeprilysin) (Catalog # 1182-ZNC)
- Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005), Prepare 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhNeprilysin to 0.1 µg/mL in Assay Buffer.
- Dilute fluorogenic peptide Substrate to 20 µM in Assay Buffer.
- Load in a black well plate 50 µL of 0.1 µg/mL of rhNeprilysin (5 ng/well), and start the reaction by adding 50 µL of 20 µM Substrate. As a Substrate Blank combine 50 µL of Substrate and 50 µL of Assay Buffer.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
- 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:
- rhNeprilysin: 0.005 µg
- Substrate: 10 µM
