Cathepsin C (CTSC), also known as
dipeptidyl-peptidase I (DPPI), is a chloride-dependent cysteine protease in the
papain family (1-3). It sequentially removes dipeptides from the free N-terminus
of proteins and peptides. It has broad specificity except that it does not
cleave proteins with a basic amino acid (Arg or Lys) in the N-terminal position
or with a proline on either side of the scissile bond. Cathepsin C is
synthesized with a large propeptide followed by a mature region, which is
further cleaved into heavy and light chains during processing. The N-terminal
region of the propeptide is known as the exclusion domain. It is present in the
mature, active enzyme and regulates access of substrates to the active site (4).
Cathepsin C is widely expressed and plays a major role in lysosomal
degradation and enzyme activation. It activates granule serine proteases in
cytotoxic T lymphocytes and natural killer cells (Granzymes A and B),
mast cells (Tryptase and Chymase), and neutrophils (Cathepsin G and Elastase)
by removing their N-terminal activation dipeptides (5).
Recombinant Rat Cathepsin C/DPPI Protein, CF
R&D Systems | Catalog # 8285-CY
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Key Product Details
- R&D Systems NS0-derived Recombinant Rat Cathepsin C/DPPI Protein (8285-CY)
- 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 rat Cathepsin C/DPPI protein
Asp25-Leu462, with a C-terminal 10-His tag
Asp25-Leu462, with a C-terminal 10-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Level
<0.1 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Asp25
Predicted Molecular Mass
51 kDa
SDS-PAGE
56-62 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Gly-Arg-7-amido-4-methylcoumarin (GR-AMC).
The specific activity is >25,000 pmol/min/μg, as measured under the described conditions.
The specific activity is >25,000 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
8285-CY
| Formulation | Supplied as a 0.2 μm filtered solution in MES 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 C/DPPI
References
- Turk, B.E. et al. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) pp. 1192, Elsevier Academic Press, San Diego.
- Minarowska, A. et al. (2012) Folia Histochem. Cytobiol. 50:20.
- Ishidoh, K. et al. (1991) J. Biol. Chem. 266:16312.
- Turk, D. et al. (2001) EMBO J. 20:6570.
- Dahl, S.W. et al. (2001) Biochemistry 40:1671.
Alternate Names
CTSC, DPPI, PALS, PLS
Gene Symbol
CTSC
UniProt
Additional Cathepsin C/DPPI Products
Product Documents for Recombinant Rat Cathepsin C/DPPI 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 Rat Cathepsin C/DPPI Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Rat Cathepsin C/DPPI Protein, CF
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Protocols
View specific protocols for Recombinant Rat Cathepsin C/DPPI Protein, CF (8285-CY):
Materials
- Activation Buffer: 25 mM MES, 5 mM DTT, pH 5.5
- Assay Buffer: 25 mM MES, 50 mM NaCl, 5 mM DTT, pH 6.0
- Recombinant Rat Cathepsin C/DPPI (rrCathepsin C) (Catalog # 8285-CY)
- Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
- Substrate: Gly-Arg-AMC (Bachem, Catalog # I-1215)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rrCathepsin C to 20 µg/mL in Activation Buffer.
- Dilute rhCathepsin L to 2 µg/mL in Activation Buffer.
- Combine equal volumes of diluted rrCathepsin C and diluted rhCathepsin L for final concentrations of 10 µg/mL and 1 µg/mL respectively.
- Incubate at room temperature for 30 minutes to activate rrCathepsin C.
- Dilute activated rrCathepsin C to 0.02 µg/mL in Assay Buffer.
- Dilute Substrate to 100 µM in Assay Buffer.
- Load 50 µL of the 0.02 µg/mL rrCathepsin C into a plate, and start the reaction by adding 50 µL of 100 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 100 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 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 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).
Per Well:
- rrCathepsin C: 0.001 µg
- Substrate: 50 µM