Spinesin, encoded by the TMPRSS5 gene, is a member of type II transmembrane serine proteases (TTSPs) (1). Human Spinesin contains the following structural domains: a short N-terminal cytoplasmic tail (amino acid residues 1-49), a transmembrane domain (residues 50-70), a stem region and a serine protease domain (residues 71-457) (2). The domain structure of Spinesin is common to other TTSPs, many of which have additional domains. The stem region of Spinesin contains a scavenger receptor-like domain. The ectodomain of human Spinesin (residues 71-457) was expressed and purified as a single chain pro-enzyme. The deduced amino acid sequence contains a Leu instead of a Phe residue at position 369; the former is identical to the mouse protein (3, 4). The pro-enzyme can be activated and the resulting enzyme activity can be measured as described in the Activity Assay Protocol.
Recombinant Human Spinesin Protein, CF
R&D Systems | Catalog # 2495-SE
Key Product Details
- R&D Systems NS0-derived Recombinant Human Spinesin Protein (2495-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
Structure / Form
Applications
Product Specifications
Source
Tyr71-Leu457 (Phe369Leu), with an N-terminal 9-His tag
Accession # NP_110397
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
Measured by its ability to cleave the fluorogenic peptide substrate Boc-QAR-AMC (Catalog # ES014).
The specific activity is >300 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2495-SE
| Formulation | Supplied as a 0.2 μm filtered solution in MES and NaCl. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Spinesin
References
- Hooper, J.D. et al. (2001) J. Biol. Chem. 276:857.
- Yamaguchi, Y. et al. (2002) J. Biol. Chem. 277:6806.
- Carninci, P. et al. (2000) Genome Res. 10:1617.
- Shibata, K. et al. (2000) Genome Res. 10:1757.
Alternate Names
Gene Symbol
UniProt
Additional Spinesin Products
Product Documents for Recombinant Human Spinesin 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 Spinesin Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Spinesin Protein, CF
Powered by Bioz
Customer Reviews for Recombinant Human Spinesin Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Spinesin Protein, CF and earn rewards!
Have you used Recombinant Human Spinesin Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Protocols
View specific protocols for Recombinant Human Spinesin Protein, CF (2495-SE):
- 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, pH 8.0
- Recombinant Human Spinesin (rhSpinesin) (Catalog # 2495-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
- Substrate: BOC-Gln-Ala-Arg-AMC (Catalog # ES014), 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 rhSpinesin to 200 µg/mL in Activation Buffer.
- Dilute Thermolysin to 2 µg/mL in Activation Buffer.
- Combine equal amounts of 200 µg/mL rhSpinesin and 2 µg/mL Thermolysin.
- Incubate the reactions at 37 °C for 1 hour.
- Stop the Thermolysin activity by adding 1,10 Phenanthroline to a final concentration of 10 mM.
- Dilute activated rhSpinesin to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load 50 µL of 2 ng/µL rhSpinesin into a plate, 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 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:
- rhSpinesin: 0.100 µg
- Thermolysin: 0.001 µg
- Substrate: 100 µM