Cathepsin A/lyososomal carboxypeptidase A is a member of the serine carboxypeptidase family (1). Cathepsin A, also known as protective protein, is synthesized as a single-chain precursor and processed into heavy (32 kDa) and light (20 kDa) chains, which are linked by disulfide bonds. Cathepsin A is a multifunctional enzyme that expresses deaminidase and esterase activities at neutral pH and carboxypeptidase activity at acidic pH (2, 3). Cathepsin A is capable of hydrolyzing a variety of bioactive peptide hormones including endothelin and bradykinin making it a promising target in heart failure (2-5). A hereditary variant of cathepsin A results in cathepsin A-related arteriopathy with strokes and leukoencephalapathy (CARASAL) (6). In addition, Cathepsin A's association with beta -galactosidase ( beta -gal) and neuraminidase in a complex is essential for beta -gal stability and neuraminidase activation in the lysosomes. Inherited deficiency of Cathepsin A causes Galactosialidosis, a lysosomal storage disorder, characterized by a combined secondary deficiency of beta -gal and neuraminidase (7, 8).
Recombinant Mouse Cathepsin A/Lysosom Carboxypeptidase A, CF
R&D Systems | Catalog # 9789-SE
Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Cathepsin A/Lysosom Carboxypeptidase A (9789-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
Accession Number
Applications
Product Specifications
Source
Ala24-Tyr474, with a C-terminal 10-His tag
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >160 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
9789-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: Cathepsin A/Lysosomal Carboxypeptidase A
References
- Pshezhetsky, A.V. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) p. 1923, Academic Press, San Diego.
- Hiraiwa, M. (1999) Cell. Mol. Life. Sci. 56:894.
- Schreuder, H.A. et al. (2014) Biochem. Biophys. Res. Commun. 445:451.
- Ruf, S. et al. (2013) Future Med. Chem. 5:399.
- Timur, Z.K. et al. (2016) Front. Mol. Biosci. 3:68.
- Bugiani, M. et al. (2016) Neurology 84:1777.
- Caciotti, A. et al. (2013) Ophanet. J. Rare Dis. 8:114.
- Annunziata, I. et al. (2017) Expert Opin. Orphan Drugs 5:131.
Alternate Names
Gene Symbol
UniProt
Additional Cathepsin A/Lysosomal Carboxypeptidase A Products
- All Products for Cathepsin A/Lysosomal Carboxypeptidase A
- Cathepsin A/Lysosomal Carboxypeptidase A cDNA Clones
- Cathepsin A/Lysosomal Carboxypeptidase A ELISA Kits
- Cathepsin A/Lysosomal Carboxypeptidase A Lysates
- Cathepsin A/Lysosomal Carboxypeptidase A Primary Antibodies
- Cathepsin A/Lysosomal Carboxypeptidase A Proteins and Enzymes
Product Documents for Recombinant Mouse Cathepsin A/Lysosom Carboxypeptidase A, 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 Cathepsin A/Lysosom Carboxypeptidase A, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Cathepsin A/Lysosom Carboxypeptidase A, CF (9789-SE):
- Assay Buffer: 25 mM MES, 0.5 mM TCEP, pH 5.5
- Recombinant Mouse Cathepsin A/Lysosomal Carboxypeptidase A (rmCathepsin A) (Catalog # 9789-SE)
- Activator: Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
- E 64 (Tocris, Catalog # 5208), 50 mM stock in DMSO
- Fluorogenic Peptide Substrate V: MCA-RPPGFSAFK-(DNP)-OH (Catalog # ES005), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rmCathepsin A to 100 μg/mL in Assay Buffer.
- Dilute rhCathepsin L to 10 μg/mL in Assay Buffer.
- Combine equal volumes of rmCathepsin A and rhCathepsin L for final concentrations of 50 μg/mL and 5 μg/mL, respectively. Include a control containing Assay Buffer in place of rmCathepsin A (Cathepsin L Control).
- Incubate at room temperature for 30 minutes.
- Dilute E 64 to 20 μM in Assay Buffer.
- Combine equal volumes of activated rmCathepsin A and 20 μM E 64 to stop the reaction. Combine equal volumes of Cathepsin L Control and 20 μM E 64.
- Dilute activated rmCathepsin A to 3 μg/mL in Assay Buffer. Perform equivalent dilution on Cathepsin L Control.
- Dilute Substrate to 40 μM in Assay Buffer.
- Load into a plate 50 μL of 3 μg/mL rmCathepsin A and Cathepsin L Control, and start the reaction by adding 50 μL of 40 μM Substrate.
- Read plate at excitation and emission wavelengths of 320 nm and 405 nm, respectively, (top read) 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 Cathepsin L Control.
**Derived using calibration standard Mca-PL-OH (Bachem, Catalog # M-1975).
Per Well:
- rmCathepsin A: 0.15 μg
- Substrate: 20 μM
FAQs for Recombinant Mouse Cathepsin A/Lysosom Carboxypeptidase A, CF
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Q: Where does Cathepsin A cleave Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate, Catalog # ES005?
A: Although the QC assay conditions provided on our recombinant Cathepsin A datasheets should favor carboxypeptidase activity, it is possible that there is more than one site in the ES005 peptide recognized and cleaved by Cathepsin A. We have not performed verification experiments to confirm the cleavage site preferred in our reaction conditions.