Recombinant Human Enteropeptidase/Enterokinase Protein, CF

R&D Systems | Catalog # 10438-SE

Activated
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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Enteropeptidase/Enterokinase Protein (10438-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

P98073

Structure / Form

Activated, heavy and light chains

Applications

Enzyme Activity
View all Enteropeptidase/Enterokinase Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Enteropeptidase/Enterokinase protein
Leu41-His1019
with a C-terminal 9-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

N-terminal Sequence Analysis

Leu178, Ala294, Ile306 (heavy chain) & Ile785 (light chain)

Predicted Molecular Mass

67, 55, 53 kDa (heavy chain) & 27 kDa (light chain)

SDS-PAGE

98-117 kDa (heavy chain) & 46-49 kDa (light chain), reducing conditions

Activity

Measured by its ability to cleave a colorimetric peptide substrate, Z-Lys-SBzl.
The specific activity is >30,000 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human Enteropeptidase/Enterokinase Protein, CF

Recombinant Human Enteropeptidase/Enterokinase Protein Enzyme Activity

Recombinant Human Enteropeptidase/Enterokinase Protein Enzyme Activity

Recombinant Human Enteropeptidase/Enterokinase (Catalog # 10438-SE) is measured by its ability to cleave a colorimetric peptide substrate, Z-Lys-SBzl.
Recombinant Human Enteropeptidase/Enterokinase Protein SDS-PAGE

Recombinant Human Enteropeptidase/Enterokinase Protein SDS-PAGE

2 μg/lane of Recombinant Human Enteropeptidase/Enterokinase (Activated) (Catalog # 10438-SE) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at approximately 110, 100, & 47 kDa under reducing conditions.

Formulation, Preparation, and Storage

10438-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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Enteropeptidase/Enterokinase

Enteropeptidase, also known as enterokinase, is a type II transmembrane serine protease that initiates activation of pancreatic proteases by converting trypsinogen to trypsin, which subsequently leads to activation of chymotrypsin, carboxypeptidases and elastases in the intestine (1). Located in the intestinal brush border, enteropeptidase is a disulfide bond-linked dimer of an N-terminal heavy chain (HC) and C-terminal light chain (LC) derived from the same single-chain precursor. The non-catalytic multidomain HC includes a short cytoplasmic tail, a transmembrane, a MSCR, a MAM, two CUB, and two LDL-receptor class A domains while the LC contains the catalytic domain of trypsin-like serine proteases (1,2). Enteropeptidase is known to have high sequence specificity making it useful as a biotechnological tool for recombinant fusion domains. Human enteropeptidase LC has greater efficiency and specificity than bovine enteropeptidase LC (3,4). Removal of HC domains results in significant loss of activity towards its physiological substrate trypsinogen (5-7) although cleavage of small peptidyl substrates remains equivalent (6,8). Enteropeptidase inhibition may be a target in diabetes and obesity (9,10). The purified activated recombinant human Enteropeptidase corresponds to the heterodimer of LC and HC without the transmembrane domain.

References

  1. Zheng, X.L. et al. (2009) Front. Biosci. 1:242.
  2. Lu, D. et al. (1999) J. Mol. Biol. 292:361.
  3. Gasparian, M.E. et al. (2006) Biochemistry 71:113.
  4. Mikhailova, A.G. et al. (2007) Protein Pept. Lett. 14:227.
  5. LaVallie, E.R. et al. (1993) J. Biol. Chem. 268:23311.
  6. Lu, D. et al. (1997) J. Biol. Chem. 272:31293.
  7. Mikhailova, A.G. et al. (1999) FEBS Lett. 442:226.
  8. Light, A and P Fonseca. (1984) J. Biol. Chem. 259:13195.
  9. Braud, S. et al. (2012) PLoS One 7:e49612.
  10. Yashiro, H. et al. (2019) Diabetes Obes. Metab. 21:2228.

Alternate Names

Enterokinase, ENTK, PRSS7, TMPRSS15

Entrez Gene IDs

5651 (Human)

Gene Symbol

TMPRSS15

UniProt

P98073

Additional Enteropeptidase/Enterokinase Products

Product Documents for Recombinant Human Enteropeptidase/Enterokinase Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Enteropeptidase/Enterokinase Protein, CF

For research use only

Related Research Areas

Serine Proteases and Regulators

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Protocols

View specific protocols for Recombinant Human Enteropeptidase/Enterokinase Protein, CF (10438-SE):

Materials
  •  Assay Buffer: 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% Brij-35, pH 7.5 (TCNB)
  • Recombinant Human Enteropeptidase/Enterokinase, active (rhEnterokinase) (Catalog # 10438-SE)
  • Substrate: thiobenzyl benzyloxycarbonyl-L-lysinate (Z-Lys-SBzl) (Bachem, Catalog # M-1300), 10 mM stock in DMSO
  • 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D8130), 10 mM stock in DMSO
  • 96-well clear plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhEnterokinase to 0.04 µg/mL in Assay Buffer.
  2. Dilute Substrate to 400 µM in Assay Buffer containing 400 µM of DTNB.
  3. In a plate, load 50 µL of the diluted rhEnterokinase, and start the reaction by adding 50 µL of Substrate/DTNB mixture.  Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate/DTNB mixture.
  4. Read at an absorbance of 405 nm in kinetic mode of 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)


*Adjusted for Substrate Blank
**Using the extinction coefficient 13260 M-1cm-1
***Using the path correction 0.32 cm
Note: the output of many spectrophotometers is in mOD

Per Well:

  • rhEnterokinase: 0.002 µg
  • DTNB: 200 µM
  • Substrate: 200 µM

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