Recombinant Active Human Hepsin His-tag Protein, CF

Catalog #: 11416-SE Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
11416-SE-020

Recombinant Active Human Hepsin His-tag Protein Enzyme Activity.

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Active Human Hepsin His-tag Protein, CF Summary

Product Specifications

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.

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.

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.

Accession #

P05981.1

N-terminal Sequence
Analysis

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

Structure / Form

Active form

Predicted Molecular Mass

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

SDS-PAGE

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

Product Datasheets

11416-SE

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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

Dilute rhHepsin, active to 0.01 µg/mL in Assay Buffer.
Dilute Substrate to 400 µM in Assay Buffer.
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.
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 V_max* (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

Scientific Data

Enzyme Activity

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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).

Reconstitution Calculator

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

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

Entrez Gene IDs

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

Alternate Names

EC 3.4.21; EC 3.4.21.106; Hepsin; HPN; serine protease hepsin catalytic chain; serine protease hepsin; TMPRSS1 ; TMPRSS1serine protease hepsin non-catalytic chain; Transmembrane protease serine 1; transmembrane protease, serine 1