ECE-2 is a zinc protease of the neprilysin (NEP) family, which also includes ECE-1, PEX, XCE, DINE, Kell and several NEP-like proteins (1). ECE-2 is a type II transmembrane protein with a short cytoplasmic tail and a large ectodomain. Three alternatively spliced forms differ in their cytoplasmic tail (2). In addition to big endothelin-1, ECE-2 cleaves a variety of bioactive peptides such as bradykinin, neurotensin, angiotensin I, substance P, dynorphin B, proenkephalin-derived peptides such as peptide E, BAM 22 and BAM18, and PEN-LEN, an endogenous inhibitor of prohormone convertase 1 (3). Together with ECE-1, it is also involved in degradation of beta -amyloid peptide (4). The ectodomain of human ECE-2 was expressed with an N-terminal His tag and purified.
Recombinant Human ECE-2 Protein, CF
R&D Systems | Catalog # 1645-ZN
Key Product Details
- R&D Systems NS0-derived Recombinant Human ECE-2 Protein (1645-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
Accession Number
Applications
Product Specifications
Source
Gly104-Trp787, with an N-terminal 10-His tag and Val
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005).
The specific activity is > 250 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1645-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and ZnCl2. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: ECE-2
References
- Turner, A.J. et al. (2001) BioEssays 23:261.
- Lorenzo, M.N. et al. (2001) Biochim. Biophys. Acta 1522:46.
- Mzhavia, N. et al. (2003) J. Biol. Chem. 278:14704.
- Eckman, E.A. et al. (2003) J. Biol. Chem. 278:2081.
Long Name
Alternate Names
Entrez Gene IDs
Gene Symbol
UniProt
Additional ECE-2 Products
Product Documents for Recombinant Human ECE-2 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 ECE-2 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ECE-2 Protein, CF
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Protocols
View specific protocols for Recombinant Human ECE-2 Protein, CF (1645-ZN):
- Assay Buffer: 0.1 M MES, 0.25 M NaCl, pH 5.75
- Recombinant Human ECE-2 (rhECE-2) (Catalog # 1645-ZN)
- Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhECE-2 to 100 µg/mL in Assay Buffer.
- Incubate at room temperature for 30 minutes.
- Dilute the incubated rhECE-2 to 0.8 ng/µL.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of 0.8 ng/µL rhECE-2 into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing Assay Buffer and 20 µM Substrate.
- 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:
- rhECE-2: 0.040 µg
- Substrate: 10 µM
Associated Pathways
