Hepatocyte Growth Factor Activator (HGFA) is a serine endopeptidase that cleaves at the peptide bond between Arg494 and Val495 of single-chain human HGF precursor, generating the active heterodimer (1). HGFA is produced and secreted by the liver and normally circulates in the blood as an inactive zymogen (2, 3). The zymogen has a weak affinity for heparin but acquires a strong affinity for heparin upon activation that is linked to blood coagulation. This property may ensure the local action of this enzyme at the site of tissue injury (3). Human HGFA precursor (655 amino acid residues) contains several predicted domains including a signal peptide (residues 1‑30), a propeptide (residues 31-372), and a mature and active form (residues 373-655) that is further processed into a short chain (residues 373‑407) and a long chain (residues 408-655). The short chain and the long chain (catalytic domain) may form a disulfide bond linked dimer. HGFA can be activated by thrombin (Catalog # http://www.rndsystems.com/product_results.aspx?k=1473-SE">1473-SE) or thermolysin (Catalog # http://www.rndsystems.com/product_results.aspx?k=3097-ZN">3097-ZN) (4). The active protease can be inhibited by HGFA inhibitors (HAIs). Two HAIs, HAI-1 and HAI-2, are known in mouse and human. HAI-1 is not only an inhibitor, but also a specific acceptor of active HGFA, acting as a reservoir of this enzyme on the cell surface (5).
Recombinant Human HGF Activator Protein, CF
R&D Systems | Catalog # 1514-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Human HGF Activator Protein (1514-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 HGF Activator protein
Gln36-Ser655, with a C-terminal 10-His tag
Gln36-Ser655, 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
Gln36
Predicted Molecular Mass
68 kDa
SDS-PAGE
105 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 >15 pmol/min/µg, as measured under the described conditions.
The specific activity is >15 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1514-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2. |
| 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: HGF Activator
References
- Kitamura, N. (2004) in Handbook of Proteolytic Enzymes (Barrett, A.J. et al. Eds.) p. 1712, Academic Press, San Diego.
- Miyazawa, K. et al. (1993) J. Biol. Chem. 268:10024.
- Miyazawa, K. et al. (1996) J. Biol. Chem. 271:3615.
- Shimomura, T. et al. (1993) J. Biol. Chem. 268:22927.
- Kataoka, H. et al. (2000) J. Biol. Chem. 275:40453.
Long Name
Hepatocyte Growth Factor Activator
Alternate Names
HGFA, Hgfac
Gene Symbol
HGFAC
UniProt
Additional HGF Activator Products
Product Documents for Recombinant Human HGF Activator 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 HGF Activator Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human HGF Activator Protein, CF
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Protocols
View specific protocols for Recombinant Human HGF Activator Protein, CF (1514-SE):
Materials
- Activation Buffer: 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 10 µg/mL dextran sulfate, pH 7.5
- Assay Buffer: 50 mM Tris, 0.01% (w/v) CHAPS, pH 9.0
- Recombinant Human HGF Activator (rhHGFA) (Catalog # 1514-SE)
- Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
- 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
- Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhHGFA to 100 µg/mL in Activation Buffer containing Thermolysin at 4.3 µg/mL.
- Incubate at 37 °C for 1 hour.
- Dilute 1,10 Phenanthroline to 20 mM in Assay Buffer.
- Add an equal volume of 20 mM 1,10 Phenanthroline (final concentration 10 mM) to stop Thermolysin activity.
- Incubate at room temperature for 30 minutes.
- Dilute rhHGFA to 10 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load in a black well plate 50 µL of 10 ng/µL rhHGFA, and start the reaction by adding 50 µL of 20 µM Substrate. Prepare a Substrate Blank by combining 50 µL of 20 µM Substrate with 50 µL of Assay Buffer.
- Read at excitation and emission wavelengths of 320 nm and 405 nm, 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:
- rhHGFA: 0.5 µg
- Substrate: 10 µM
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