Human Trypsin 3, encoded by the PRSS3 gene, is also known as mesotrypsin (1). Constituting less than 10% of the total trypsin content in normal pancreatic juice, it is one of the three trypsin isoforms produced by the pancreas (2). Compared to Trypsin 1 and 2, one intriguing feature of Trypsin 3 is its resistance to polypeptide trypsin inhibitors, such as the Kunitz-type soybean trypsin inhibitor or the Kazal-type pancreatic secretory trypsin inhibitor. As revealed by the crystal structure, this resistance is likely due to the presence of an arginine residue in place of the highly conserved Gly198 (3). Trypsin 3 is synthesized in the pancreas and secreted into the duodenum lumen, where it is activated by enterokinase. One physiologic function of Trypsin 3 has been proposed to be degradation of trypsin inhibitors, which facilitates the digestion of those foods rich in these proteins (4).
Recombinant Human Trypsin 3/PRSS3 Protein, CF
R&D Systems | Catalog # 3710-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Trypsin 3/PRSS3 Protein (3710-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 human Trypsin 3/PRSS3 protein
Val16-Ser247, with a C-terminal 10-His tag
Val16-Ser247, with a C-terminal 10-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Val16
Predicted Molecular Mass
27 kDa
SDS-PAGE
32 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002).
The specific activity is >4,000 pmol/min/μg, as measured under the described conditions.
The specific activity is >4,000 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
3710-SE
| 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: Trypsin 3/PRSS3
References
- Nyaruhucha, C.N.M. et al. (1997) J. Biol. Chem. 272:10573.
- Rinderknecht, H. et al. (1984) Gastroenterology 86:681.
- Katona, G. et al. (2002) J. Mol. Biol. 315:1209.
- Szmola, R. et al. (2003) J. Biol. Chem. 278:48580.
Alternate Names
Mesotrypsin, MTG, PRSS3, PRSS4, Trypsin 4, Trypsinogen 15
Gene Symbol
PRSS3
UniProt
Additional Trypsin 3/PRSS3 Products
Product Documents for Recombinant Human Trypsin 3/PRSS3 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 Trypsin 3/PRSS3 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Trypsin 3/PRSS3 Protein, CF
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Protocols
View specific protocols for Recombinant Human Trypsin 3/PRSS3 Protein, CF (3710-SE):
Materials
- Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 8.0
- Recombinant Human Trypsin 3/PRSS3 (rhTrypsin 3) (Catalog # 3710-SE)
- Recombinant Human Enteropeptidase/Enterokinase (rhEnterokinase) (Catalog # 1585-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- Substrate: Mca-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(Dnp)-NH2 (Catalog # ES002)
- 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Activate rhEnterokinase with Thermolysin.
- Dilute rhEnterokinase to 100 µg/mL with Activation Buffer.
- Dilute Thermolysin to 3.16 μg/mL with Activation Buffer.
- Mix equal volumes of diluted rhEnterokinase with Thermolysin
- Incubate at 37 °C for 30 minutes.
- Stop the reaction by adding an equal volume of 20 mM 1,10 Phenanthroline.
- Activate rhTrypsin 3, with activated rhEnterokinase.
- Dilute the activated rhEnterokinase to 2 µg/mL in Assay Buffer.
- Dilute rhTrypsin 3 to 200 µg/mL in Assay Buffer.
- Mix equal volumes of the diluted rhEnterokinase and diluted rhTrypsin 3.
- Incubate at 37 °C for 90 minutes.
- After incubation, dilute activated rhTrypsin 3 to 0.1 µg/mL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load in a black well plate 50 µL of rhTrypsin 3 at 0.1 µg/mL, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of 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:
- rhTrypsin 3: 0.005 µg
- Substrate: 10 µM
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