Recombinant Active Human Hepsin His-tag Protein, CF

R&D Systems | Catalog # 11416-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Active Human Hepsin His-tag Protein (11416-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

P05981.1

Structure / Form

Active form

Applications

Enzyme Activity
View all Hepsin Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Hepsin protein
Arg45 to Leu417 (Asp161Glu, Arg162Lys) with a C-terminal 10-His tag
The proform was activated.

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Arg45 (non-catalytic chain) & Ile163 (catalytic chain)

Predicted Molecular Mass

13 kDa (non-catalytic chain), 29 kDa (catalytic chain)

SDS-PAGE

17-24 and 29-33 kDa, under reducing conditions

Activity

Measured by its ability to cleave tert-butoxycarbonyl-Gln-Arg-Arg-7-amino-4-methylcoumarin (Boc-QRR-AMC).
The specific activity is >45,000 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Active Human Hepsin His-tag Protein, CF

Recombinant Active Human Hepsin His-tag Protein Enzyme Activity.

Recombinant Active Human Hepsin His-tag Protein (Catalog # 11416-SE) is measured by its ability to cleave tert-butoxycarbonyl-Gln-Arg-Arg-7-amino-4-methylcoumarin (Boc-QRR-AMC).

Formulation, Preparation, and Storage

11416-SE
Formulation Supplied as a 0.2 μm filtered solution in Tris 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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Hepsin

Hepsin, also known as TMPRSS1, is part of a family of type II transmembrane serine proteases (TTSPs) (1) that cleaves critical target proteins including Hepatocyte growth factor (HGF), uromodulin (UMOD), uPA, and macrophage-stimulating protein (MSP) at specific cleavage sites following basic residues with a strong preference for arginine residues (2-5). Hepsin is a 417 amino acid polypeptide that is activated to a two-chain disulfide-linked form. Hepsin, like other proteases in this family, contains an N-terminal cytoplasmic tail and transmembrane domain, and an extracellular stem region with an SRCR domain and C-terminal serine protease domain with a conserved activation site (1, 6). It is most highly expressed in liver, but is also present in many other tissues, notably lung, kidney, and skeletal muscle (7) and is found at high levels in several cancers including prostate, breast, and ovarian (2, 5, 8-10). In the liver, hepsin functions to regulate glucose, lipid, and protein metabolism through proteolytic conversion of pro-HGF to HGF, a potent ligand for Met signaling and dysregulation (11). Involvement in cancers has made hepsin a target of interest using either small molecule inhibitors or antibodies to neutralize their proteolytic activity, for example for prostate cancer and metastasis (12-14). Hepsin is also of interest for its potential role in osteoarthritis (15) and its ability to cleave surface proteins of respiratory viruses making it a therapeutic target for respiratory viral diseases (13). Recombinant Human Hepsin was expressed as a secreted, soluble protein lacking its cytosolic and transmembrane domains and activated. 

References

  1. Li. S. et al. (2021) FEBS J. 288:5252.
  2. Herter, S. et al. (2005) Biochem. J. 390:125.
  3. Ganesan, R. et al. (2011) Mol. Cancer Res. 9:1175.
  4. Brunati, M. et al. (2015) Elife 4:e08887.
  5. Tanabe, L.M. and K. List (2017) FEBS J. 284:1421.
  6. Somoza, J.R. et al. (2003) Structure 11:1123.
  7. Tsuji, A. et al. (1991) J. Biol. Chem. 266:16948.
  8. Dhanasekaran, S.M. et al. (2001) Nature 412:822.
  9. Miao, J. et al. (2008) Int. J. Cancer 123:2041.
  10. Xing, P. et. al. (2011) J. Investig. Med. 59:803.
  11. Li, S. et al. (2020) Proc. Natl. Acad. Sci. USA 117:12359.
  12. Damalanka, V.C. et al. (2019) J. Med. Chem. 62:480.
  13. Murza, A. et al. (2020) Expert Opin. Ther. Pat. 30:807.
  14. Wu, Q. and Parry, G. (2007) Front. Biosci. 12:5052.
  15. Wilkinson, D.J. et al. (2017) Sci. Rep. 7:16693.

Alternate Names

HPN, TMPRSS1

Entrez Gene IDs

3249 (Human); 15451 (Mouse); 29135 (Rat)

Gene Symbol

HPN

UniProt

P05981.1

Additional Hepsin Products

Product Documents for Recombinant Active Human Hepsin His-tag Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Active Human Hepsin His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Active Human Hepsin His-tag Protein, CF (11416-SE):

Materials
  • Assay Buffer:  50 mM Tris, 0.05% Brij-35, pH 9.0
  • Recombinant Active Human HepsinHis-tag (rhHepsin) (Catalog # 11416-SE)
  • Substrate:  Boc-QRR-AMC, 5 mM stock in DMSO
  • Black 96-well Plate
  • Plate Reader with Fluorescence Read Capability
  1. Dilute rhHepsin, active to 0.01 µg/mL in Assay Buffer.
  2. Dilute Substrate to 400 µM in Assay Buffer.
  3. Load in a plate 50 μL of 0.01 µg/mL rhHepsin, active, and start the reaction by adding 50 μL of 400 μM Substrate. Include a Substrate Blank containing 50 μL of 400 μM Substrate and 50 μL of Assay Buffer.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. 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
Per Well
  • rhHepsin, active:  0.0005 μg
  • Substrate:  200 µM






















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