Mast cell protease-1 (Mcpt1), also known as beta ‑chymase, is a member of the Chymase family of chymotrypsin-like serine proteases (1). mMcpt1 is a mast cell protease predominantly expressed in intestinal mucosal mast cells where it promotes mucosal permeability in intestinal allergic hypersensitivity reactions (2). Its activation is completed by the removal of a two residue N‑terminal propeptide by a dipeptidyl peptidase (Cathepsin C) (3). Like human alpha ‑Chymase, Mcpt1 is capable of the conversion of angiotensin I to angiotensin II, which plays a key role in the regulation of arterial pressure (4). Studies have shown that specific chymase inhibitors are able to diminish the development of abdominal aortic aneurysm and reduce the adhesion formation after cardiac surgery in hamsters (5, 6). Therefore, the development of specific inhibitors of chymase activity has been a pharmacologic strategy to develop therapeutic agents.
Recombinant Mouse Mast Cell Protease-1/Mcpt1 Protein, CF
R&D Systems | Catalog # 5146-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Mast Cell Protease-1/Mcpt1 Protein (5146-SE)
- 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
Structure / Form
Pro form
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Mast Cell Protease-1/Mcpt1 protein
Glu19-Lys246, with a C-terminal 10-His tag
Glu19-Lys246, with a C-terminal 10-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
Glu19
Predicted Molecular Mass
27 kDa
SDS-PAGE
33 kDa, 35 kDa and 36 kDa, reducing conditions
Activity
Measured by its ability to cleave Suc-Ala-Ala-Pro-Phe-AMC (Suc-AAPF-AMC, Bachem, Catalog # I-1465).
The specific activity is >35 pmol/min/µg, as measured under the described conditions.
The specific activity is >35 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
5146-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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: Mast Cell Protease-1/Mcpt1
References
- Caughey, G.H. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.), p. 1531 Academic Press, San Diego.
- Wastling, J.M. et al. (1998) Am. J. Pathol. 153:491.
- Murakami, M. et al. (1995) J. Biol. Chem. 270:2218.
- Saito, K. et al. (2003) Biochem. Biophys. Res. Commun. 302:773.
- Tsunemi, K. et al. (2004) J. Pharmacol. Exp. Ther. 309:879.
- Soga, Y. et al. (2004) J. Thorac. Cardiovasc. Surg. 127:72.
Alternate Names
Mast Cell Protease1, Mcp1, MMPC-1
Gene Symbol
MCPT1
UniProt
Additional Mast Cell Protease-1/Mcpt1 Products
Product Documents for Recombinant Mouse Mast Cell Protease-1/Mcpt1 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 Mouse Mast Cell Protease-1/Mcpt1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse Mast Cell Protease-1/Mcpt1 Protein, CF
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Protocols
View specific protocols for Recombinant Mouse Mast Cell Protease-1/Mcpt1 Protein, CF (5146-SE):
Materials
- Activation Buffer: 50 mM MES, 50 mM NaCl, 5 mM DTT, pH 5.5
- Assay Buffer: 50 mM Tris, 2 M NaCl, 0.025% (w/v) Brij-35, pH 8.0
- Recombinant Mouse Mast Cell Protease‑1/Mcpt1 (rmMcpt1) (Catalog # 5146-SE)
- Recombinant Mouse Active Cathepsin C/DPPI (rmCathepsin C) (Catalog # 2336-CY)
- Heparin (Sigma, Catalog # H3393), 20 mg/mL stock in deionized water
- N-Ethylmaleimide (NEM) (Sigma, Catalog # E1271), 50 mM stock in deionized water
- Substrate: SUC-Ala-Ala-Pro-Phe-AMC (Bachem, Catalog # I-1465), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmCathepsin C to 120 µg/mL in Activation Buffer.
- Dilute rmMcpt1 to 200 µg/mL in Activation Buffer.
- Dilute Heparin to 160 µg/mL in Activation Buffer.
- Combine 15 µL of 200 µg/mL rmMcpt1, 15 µL of 120 µg/mL rmCathepsin C and 30 µL of 160 µg/mL Heparin. Mix well.
- Incubate reaction mixture for 2 hours at 37 °C.
- Dilute NEM to 10 mM in Assay Buffer.
- Stop activation by adding 60 µL of 10 mM NEM.
- Dilute activated rmMcpt1 to 8 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load into a plate 50 µL of 8 ng/µL rmMcpt1, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
|
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 (AMC) (Sigma, Catalog # A-9891).
Per Well:
- rmMcpt1: 0.4 µg
- Substrate: 100 µM
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