Recombinant Human Enteropeptidase/Enterokinase Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Enteropeptidase/Enterokinase Protein, CF Summary

Details of Functionality
Measured by its ability to cleave a colorimetric peptide substrate, Z-Lys-SBzl. The specific activity is >10,000 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Enteropeptidase/Enterokinase protein
Leu41-His1019 & Ser118-His1019, with a C-terminal 9-His tag
Accession #
N-terminal Sequence
Leu41 & Ser118
Protein/Peptide Type
Recombinant Enzymes
Gene
TMPRSS15
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
110 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
150 kDa, reducing conditions
Publications
Read Publications using
1585-SE in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl2, 0.05% Brij-35, pH 7.5 (TCNB)
  • Recombinant Human Enteropeptidase/Enterokinase (rhEnterokinase) (Catalog # 1585-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
  • Substrate: thiobenzyl benzyloxycarbonyl-L-lysinate (Z-Lys-SBzl) (Bachem, Catalog # M-1300), 10 mM stock in DMSO
  • 96 well Clear Plate (Costar, Catalog #  92592)
  • 5,5’-dithio-bis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Activate rhEnterokinase with Thermolysin.
    1. Dilute rhEnterokinase to 100 μg/mL in Assay Buffer.
    2. Dilute Thermolysin to 3.16 μg/mL in Assay buffer.
    3. Mix equal volumes of diluted rhEnterokinase and Thermolysin.
    4. Incubate at 37 °C for 30 minutes.
    5. Stop the reaction by adding an equal volume of 20 mM 1,10 Phenanthroline to the reaction tube.
  2. Dilute rhEnterokinase to 0.1 μg/mL in Assay Buffer.
  3. Dilute Substrate to 200 μM in Assay Buffer with 200 μM of DTNB.
  4. Load in a 96 well clear plate 50 μL of the diluted rhEnterokinase. For a Substrate Blank load 50 μL of the Assay Buffer.
  5. Start the reaction by adding 50 μL of the Substrate/DTNB mixture to wells.
  6. Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/M
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: 0.005 μg
  • DTNB: 100 μM
  • Substrate: 100 μM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Enteropeptidase/Enterokinase Protein, CF

  • EC 3.4.21
  • EC 3.4.21.9
  • Enterokinase
  • Enteropeptidase
  • ENTK
  • ENTKenterokinase
  • MGC133046
  • protease, serine, 7 (enterokinase)
  • PRSS7
  • PRSS7enteropeptidase
  • Serine protease 7
  • TMPRSS15
  • Transmembrane protease serine 15
  • transmembrane protease, serine 15

Background

EK initiates activation of pancreatic proteases by converting trypsinogen to trypsin, which in turn activates chymotrypsin, carboxypeptidases and elastases. Located in intestinal brush border, it is a disulfide bond linked dimer of the heavy and light chains, which are derived from the same single-chain precursor. The multidomain‑containing the heavy chain consists of a short cytoplamic tail, a transmembrane, a SEA, a SRCR, a MAM, two CUB and two LDL-receptor class A domains. The light chain contains the catalytic domain of trypsin-like serine proteases. The purified recombinant human EK corresponds to the single-chain form starting at the end of the transmembrane domain.

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Publications for Enteropeptidase/Enterokinase (1585-SE)(7)

We have publications tested in 3 confirmed species: Human, Mouse, N/A.

We have publications tested in 2 applications: Bioassay, Enzyme Assay.


Filter By Application
Bioassay
(3)
Enzyme Assay
(3)
All Applications
Filter By Species
Human
(4)
Mouse
(2)
N/A
(1)
All Species
Showing Publications 1 - 7 of 7.
Publications using 1585-SE Applications Species
V Toldi, A Szabó, M Sahin-Tóth Inactivation of mesotrypsin by chymotrypsin C prevents trypsin inhibitor degradation J. Biol. Chem., 2020;0(0):. 2020 [PMID: 32014997] (Enzyme Assay, Human) Enzyme Assay Human
A Geisz, Z Jancsó, BC Németh, E Hegyi, M Sahin-Tóth Natural single-nucleotide deletion in chymotrypsinogen C gene increases severity of secretagogue-induced pancreatitis in C57BL/6 mice JCI Insight, 2019;5(0):. 2019 [PMID: 31211695] (Bioassay, Mouse) Bioassay Mouse
E Hegyi, M Sahin-Tóth Trypsinogen isoforms in the ferret pancreas Sci Rep, 2018;8(1):15094. 2018 [PMID: 30305676] (N/A) N/A
Szabo A, Radisky E, Sahin-Toth M Zymogen activation confers thermodynamic stability on a key peptide bond and protects human cationic trypsin from degradation. J Biol Chem, 2014;289(8):4753-61. 2014 [PMID: 24403079] (Bioassay, Human) Bioassay Human
Nemeth B, Wartmann T, Halangk W, Sahin-Toth M Autoactivation of mouse trypsinogens is regulated by chymotrypsin C via cleavage of the autolysis loop. J Biol Chem, 2013;288(33):24049-62. 2013 [PMID: 23814066] (Bioassay, Mouse) Bioassay Mouse
Prohaska TA, Wahlmuller FC, Furtmuller M, Geiger M Interaction of protein C inhibitor with the type II transmembrane serine protease enteropeptidase. PLoS ONE, 2012;7(6):e39262. 2012 [PMID: 22723979] (Enzyme Assay, Human) Enzyme Assay Human
Nemoda Z, Sahin-Toth M Chymotrypsin C (caldecrin) stimulates autoactivation of human cationic trypsinogen. J. Biol. Chem., 2006;281(17):11879-86. 2006 [PMID: 16505482] (Enzyme Assay, Human) Enzyme Assay Human

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Bioinformatics

Gene Symbol TMPRSS15
Uniprot