Endothelin-converting Enzyme 1 (ECE-1) is a zinc protease of the neprilysin (NEP) family, which also includes ECE-2, PEX, XCE, DINE, Kell and several NEP-like proteins (1). ECE-1 is a type II transmembrane protein with a short cytoplasmic tail and a large ectodomain. Four alternatively spliced isoforms differ in their cytoplasmic tail (2, 3). In addition to big endothelin-1, ECE-1 cleaves a variety of bioactive peptides such as bradykinin, neurotensin, angiotensin I, and substance P (1). Together with ECE-2, it is also involved in degradation of beta -amyloid peptide (4). The ectodomain of human ECE-1, which is common to all isoforms, was expressed with an N-terminal His tag and purified.
Recombinant Human ECE-1 Protein, CF
R&D Systems | Catalog # 1784-ZN
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Key Product Details
- R&D Systems NS0-derived Recombinant Human ECE-1 Protein (1784-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
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human ECE-1 protein
Gln90-Trp770, with an N-terminal signal peptide, 10-His tag and Val
Gln90-Trp770, with an N-terminal signal peptide, 10-His tag and Val
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
79 kDa
SDS-PAGE
115 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005).
The specific activity is >2,500 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1784-ZN
| 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: ECE-1
References
- Turner, A.J. et al. (2001) BioEssays 23:261.
- Valdennaire, O. et al. (1999) Eur. J. Biochem. 264:341.
- Schweizer, A. et al. (1997) Biochem. J. 328:871.
- Eckman, E.A. et al. (2003) J. Biol. Chem. 278:2081.
Long Name
Endothelin-converting Enzyme-1
Alternate Names
ECE1
Entrez Gene IDs
1889 (Human)
Gene Symbol
ECE1
UniProt
Additional ECE-1 Products
Product Documents for Recombinant Human ECE-1 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-1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ECE-1 Protein, CF
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Protocols
View specific protocols for Recombinant Human ECE-1 Protein, CF (1784-ZN):
Materials
- Assay Buffer: 0.1 M MES, 0.1 M NaCl, pH 6.0
- Recombinant Human ECE-1 (rhECE-1) (Catalog # 1784-ZN)
- Fluorogenic Peptide Substrate V: 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-1 to 0.1 µg/mL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load into a black well plate 50 µL of 0.1 µg/mL of rhECE-1, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 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-1: 0.005 μg
- Substrate: 10 µM
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Associated Pathways
