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). Mouse HGFA precursor (653 amino acid residues) contains several predicted domains including a signal peptide (residues 1‑29), a propeptide (residues 30‑369), and a mature and active form (residues 370 to 653) that is further processed into a short chain (residues 370‑405) and a long chain (residues 406‑653). The short chain and the long chain (catalytic domain) may form a disulfide bond linked dimer. HGFA can be activated by autocatalysis or by thrombin (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 Mouse HGF Activator Protein, CF
R&D Systems | Catalog # 1200-SE
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse HGF Activator Protein (1200-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
Active form
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse HGF Activator protein
Met1-Ser653 with a C-terminal 10-His tag
The pro form was purified, activated and further purified.
Met1-Ser653 with a C-terminal 10-His tag
The pro form was purified, activated and further purified.
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
Val370 & Ile406
Predicted Molecular Mass
32 kDa & 28 kDa
SDS-PAGE
35 kDa and 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 >100 pmol/min/µg, as measured under the described conditions.
The specific activity is >100 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1200-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2. |
| 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: 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 Mouse 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 Mouse HGF Activator Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse HGF Activator Protein, CF
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Protocols
View specific protocols for Recombinant Mouse HGF Activator Protein, CF (1200-SE):
Materials
- Assay Buffer: 50 mM Tris, 0.01% CHAPS, pH 9.0
- Recombinant Mouse HGFA (rmHGFA) (Catalog # 1200-SE)
- 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 rmHGFA to 2 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of 2 ng/µL rmHGFA in a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include 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:
- rmHGFA: 0.100 µg]
- Substrate: 10 µM
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