Recombinant Human Matriptase/ST14 Catalytic Domain, CF

Product Discontinued

Recombinant Human Matriptase/ST14 Catalytic Domain, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate Boc-QAR-AMC (Catalog # ES014). The specific activity is >10,000 pmol/min/µg, as measured under the described conditions.
• Source: E. coli-derived human Matriptase/ST14 protein
  Gly596-Val855, with an N-terminal Met and 6-His tag
  The protein was purified, auto-activated and further purified.
• Accession #: NP_068813
• N-terminal Sequence: Val615
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ST14
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 26 kDa; Other fragments of 19 kDa, 17 kDa, 9 kDa and 6 kDa are also observed in the recombinant protein preparation..
  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: 27 kDa (major), 20 kDa, 17 kDa, 10 kDa, 6.5 kDa, reducing conditions

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-HCl and Glycerol.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 50 mM NaCl, 0.01% (v/v) Tween® 20, pH 9.0
  • Recombinant Human Matriptase/ST14 Catalytic Domain (rhMatriptase) (Catalog # 3946-SE)
  • Substrate: BOC-Gln-Ala-Arg-AMC (Catalog # ES014), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc Cat. # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMatriptase to 0.1 µg/mL in Assay Buffer.
  2. Dilute Substrate to 50 µM in Assay Buffer.
  3. Load 50 µL of 0.1 µg/mL rhMatriptase into a plate, and start the reaction by adding 50 µL of 50 µM substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  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 (AMC) (Sigma, Cat. # A-9891).

  Per Well:
  • rhMatriptase: 0.005 µg
  • Substrate: 25 µM

Notes

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

Alternate Names for Recombinant Human Matriptase/ST14 Catalytic Domain, CF

• EC 3.4.21
• Epithin
• HAI
• Matriptase
• Membrane-type serine protease 1
• MTSP1
• MT-SP1EC 3.4.21.109
• prostamin
• PRSS14
• Serine protease 14
• Serine protease TADG-15
• SNC19
• SNC19MTSP1
• ST14
• suppression of tumorigenicity 14 (colon carcinoma)
• suppression of tumorigenicity 14 (colon carcinoma, matriptase, epithin)
• suppressor of tumorigenicity 14 protein
• TADG15
• TADG-15
• TMPRSS14
• tumor associated differentially expressed gene 15 protein
• Tumor-associated differentially-expressed gene 15 protein

Background

Human matriptase, encoded by the ST14 (suppression of tumorogenicity 14) gene, is also known as tumor associated differentially expressed gene 15 protein/TADG‑15), epithin, and membrane‑type serine protease 1/MT‑SP1 (1). Predicted to have a significant role in tumor biology, matriptase may be a novel target for anti‑cancer therapy (2). However, expressed in most human epithelia, matriptase is also important in several physiological processes (1). For example, it activates prostasin to initiate a protease cascade that is essential for epidermal differentiation (3), and it converts a single‑chain IGFBP-rp1 into the two‑chain form (4).

Matriptase is a type II transmembrane serine protease with a complex modular structure (1). The 855 amino acid (aa) sequence of human matriptase consists of a cytoplasmic tail (aa 1‑55), a transmembrane domain (aa 56‑76), and an extracellular portion (aa 77‑855). The latter contains the following domains: SEA (aa 86‑201), two CUBs (aa 214‑334 and 340‑447), four LDLRAs (aa 452‑486, 487‑523, 524‑560, and 566‑603), and a serine protease (aa 615‑855). The physiological activation of the single‑chain zymogen requires the cleavage at the SEA domain within the ER or Golgi, association with HAI-1, which facilitates the transport of the protease to the cell surface, and auto‑cleavage at QAR-V(615)VGG (1). The activated matriptase is inhibited by HAI-1, and the resulting HAI-1 complex can be shed from the cell surface (1). R&D Systems recombinant human (rh) ST14 corresponds to the catalytic domain, and is inhibited effectively by rhHAI-1 and rhHAI-2A (Catalog # 1048‑PI and 1106‑PI).

1. List, K. et al. (2006) Mol. Med. 12:1.
2. Uhland, K. (2006) Cell. Mol. Life Sci. 63:2968.
3. Netzel-Arnett, S. et al. (2006) J. Biol. Chem. 281:32941.
4. Ahmed, S. et al. (2006) FEBS J. 273:615.

Publications for Matriptase/ST14 (3946-SE)(15)

Publications using 3946-SE

• J Yang, C Tong, J Qi, X Liao, X Li, X Zhang, M Zhou, L Wang, C Ma, X Xi, T Chen, Y Gao, D Wu Engineering and Structural Insights of a Novel BBI-like Protease Inhibitor Livisin from the Frog Skin Secretion Toxins, 2022-04-12;14(4):. 2022-04-12 [PMID: 35448882] (Bioassay, Odorrana livida)
• S Kalogera, Y He, AC Bay-Jensen, T Gantzel, S Sun, T Manon-Jens, MA Karsdal, CS Thudium The activation fragment of PAR2 is elevated in serum from patients with rheumatoid arthritis and reduced in response to anti-IL6R treatment Scientific Reports, 2021-12-20;11(1):24285. 2021-12-20 [PMID: 34930943] (Bioassay, Human)
• B Howng, MB Winter, C LePage, I Popova, M Krimm, O Vasiljeva Novel Ex Vivo Zymography Approach for Assessment of Protease Activity in Tissues with Activatable Antibodies Pharmaceutics, 2021-09-02;13(9):. 2021-09-02 [PMID: 34575469] (Bioassay, Human)
• S Kumari, M Dandamudi, S Rani, E Behaeghel, G Behl, D Kent, NJ O'Reilly, O O'Donovan, P McLoughlin, L Fitzhenry Dexamethasone-Loaded Nanostructured Lipid Carriers for the Treatment of Dry Eye Disease Pharmaceutics, 2021-06-18;13(6):. 2021-06-18 [PMID: 34207223] (Bioassay, Human)
• A Iyer, TLR Humphries, EP Owens, KN Zhao, PP Masci, DW Johnson, D Nikolic-Pa, GC Gobe, DP Fairlie, DA Vesey PAR2 Activation on Human Kidney Tubular Epithelial Cells Induces Tissue Factor Synthesis, That Enhances Blood Clotting Frontiers in Physiology, 2021-03-10;12(0):615428. 2021-03-10 [PMID: 33776786] (Bioassay, Human)
• M Geiger, KG Stubenrauc, J Sam, WF Richter, G Jordan, J Eckmann, C Hage, V Nicolini, A Freimoser-, M Ritter, ME Lauer, H Stahlberg, P Ringler, J Patel, E Sullivan, S Grau-Richa, S Endres, S Kobold, P Umaña, P Brünker, C Klein Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody Nat Commun, 2020-06-24;11(1):3196. 2020-06-24 [PMID: 32581215] (Bioassay, Human)
• LM Chen, KX Chai Matriptase cleaves the amyloid-beta peptide 1-42 at Arg-5, Lys-16, and Lys-28 BMC Res Notes, 2019-01-03;12(1):5. 2019-01-03 [PMID: 30606244] (Enzyme Assay)
• T Tamberg, Z Hong, D Schepper, S Skovbjerg, DM Dupont, L Vitved, CR Schar, K Skjoedt, LK Vogel, JK Jensen Blocking the proteolytic activity of zymogen matriptase�with antibody-based inhibitors J. Biol. Chem., 2018-11-08;0(0):. 2018-11-08 [PMID: 30409910] (Enzyme Assay)
• MR Straus, GR Whittaker A peptide-based approach to evaluate the adaptability of influenza A virus to humans based on its hemagglutinin proteolytic cleavage site PLoS ONE, 2017-03-30;12(3):e0174827. 2017-03-30 [PMID: 28358853] (Enzyme Assay, Virus - Influenza A)
• A Selective Irreversible Inhibitor of Furin Does Not Prevent Pseudomonas Aeruginosa Exotoxin A-Induced Airway Epithelial Cytotoxicity PLoS ONE, 2016-07-26;11(7):e0159868. 2016-07-26 [PMID: 27459298] (Enzyme Assay, Human)
• Wood M, Cole A, Eade C, Chen L, Chai K, Cole A The HIV-1 gp41 ectodomain is cleaved by matriptase to produce a chemotactic peptide that acts through FPR2. Immunology, 2014-07-01;142(3):474-83. 2014-07-01 [PMID: 24617769] (Bioassay, Human)
• Quimbar , Pedro, Malik , Uru, Sommerhoff , Christia, Kaas , Quentin, Chan , Lai Y, Huang , Yen-Hua, Grundhuber , Maresa, Dunse , Kerry, Craik , David J, Anderson , Marilyn, Daly , Norelle High-affinity cyclic peptide matriptase inhibitors. J Biol Chem, 2013-04-02;288(19):13885-96. 2013-04-02 [PMID: 23548907] (Enzyme Assay)
• Sales KU, Masedunskas A, Bey AL Matriptase initiates activation of epidermal pro-kallikrein and disease onset in a mouse model of Netherton syndrome. Nat. Genet., 2010-07-25;42(8):676-83. 2010-07-25 [PMID: 20657595] (Bioassay, Human)
• Clark EB, Jovov B, Rooj AK Proteolytic cleavage of human acid-sensing ion channel 1 by the serine protease matriptase. J. Biol. Chem., 2010-07-02;285(35):27130-43. 2010-07-02 [PMID: 20601429] (Enzyme Assay, Xenopus)
• Myerburg MM, McKenna EE, Luke CJ, Frizzell RA, Kleyman TR, Pilewski JM Prostasin expression is regulated by airway surface liquid volume and is increased in cystic fibrosis. Am. J. Physiol. Lung Cell Mol. Physiol., 2008-02-29;294(5):L932-41. 2008-02-29 [PMID: 18310226] (Bioassay, Human)

Bioinformatics

• Gene Symbol: ST14
• Uniprot:
  • NP_068813()