Recombinant Human Proprotein Convertase 7/PCSK7 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human Proprotein Convertase 7/PCSK7 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate pERTKR-AMC (Catalog # ES013). The specific activity is >125 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Proprotein Convertase 7/PCSK7 protein
  Leu38-Thr667, with a 10-His tag between Arg141 and Ser142
• Accession #: Q16549
• N-terminal Sequence: Leu38 & His tag
• Protein/Peptide Type: Recombinant Enzymes
• Gene: PCSK7
• Purity: >95%, 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: 70 kDa (pro form) & 59 kDa (mature form).
  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: 79 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, NaCl, CaCl₂ and Glycerol.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Activation Buffer: 50 mM Tris, 10 mM CaCl₂, 0.15 M NaCl, 0.05% Brij 35, pH 7.5 (TCNB)
  • Assay Buffer: 25 mM Tris, pH 7.0
  • Recombinant Human Proprotein Convertase 7/PCSK7 (PCSK7) (Catalog # 2984-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
  • Substrate: L-PyroGlu-Arg-Thr-Lys-Arg-AMC (Catalog # ES013)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescence Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Activate rhPCSK7 by diluting to 50 µg/mL in Activation Buffer containing 2.0 µg/mL Thermolysin.
  2. Incubate activation reaction at 37 °C for 4 hours.
  3. Add 1,10 Phenanthroline at a final concentration of 10 mM to stop Thermolysin activity.
  4. Dilute Substrate to 200 µM in Assay Buffer.
  5. Diluted activated rhPCSK7 to 2.0 µg/mL in Assay Buffer.
  6. In a plate load 50 µL of 2.0 µg/mL rhPCSK7 to wells and include a Substrate Blank of 50 µL of Assay Buffer.
  7. Start the reaction by adding 50 µL of 200 µM Substrate to wells.
  8. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  9. 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 (Sigma, Catalog # A-9891).

  Per Well:
  • rhPCSK7: 0.1 µg
  • 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 Proprotein Convertase 7/PCSK7 Protein, CF

• EC 3.4.21
• EC 3.4.21.-
• EC 3.4.21.61
• EC 3.4.21.75
• EC 3.4.21.93
• hPC8
• LPC
• LPCFLJ23503
• Lymphoma proprotein convertase
• PC7FLJ45329
• PC8prohormone convertase PC7
• PCSK7
• Prohormone convertase 7
• Proprotein Convertase 7
• Proprotein convertase 8
• proprotein convertase subtilisin/kexin type 7
• SPC7
• SPC7proprotein convertase PC7
• Subtilisin/kexin-like protease PC7

Background

The human PCSK7 gene encodes proprotein convertase 1 (PC7), which is also known as PC8 and lymphoma PC (1, 2). As a serine protease of the furin/PC family, the deduced amino acid sequence of human PC7 consists of a signal peptide (residues 1 to 37), a propeptide (residue 38 to 141), and a mature chain (residues 142 to 785) that consists of extracellular (residues 142 to 667), transmembrane (residues 668 to 688) and cytoplasmic (residues 689 to 785) domains. Autocatalytic processing results in the removal of the propeptide, which confers potent inhibitory activity toward the enzyme (3-5). The purified recombinant human PCSK7 corresponds to the extracellular domain of the mature chain with a 10X His tag at the N-terminus. However, it may also contain the propeptide, as suggested by the N-terminal sequencing results. Thermolysin treatment increases its activity, which may be due to degradation of the propeptide by thermolysin.

1. Bruzzaniti, A. et al. (1996) Biochem. J. 314:727.
2. Meerabux, J. et al. (1996) Cancer Res. 56:448.
3. Van de Loo, J.-W.H.P. et al. (1997) J. Biol. Chem. 272:27116.
4. Bhattacharjya, S. et al. (2000) Biochemistry 39:2868.
5. Seidah, N.G. and M. Chretien (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 1877, Academic Press, San Diego.

Publications for Proprotein Convertase 7/PCSK7 (2984-SE)(1)

Publication using 2984-SE

• Krchlikova, V;Braun, E;Weiss, J;Stafl, K;Jech, L;Badarinarayan, SS;Lotke, R;Travnicek, M;Baur, C;Stark, P;Haussmann, I;Lu, Y;Petersen, M;Cui, W;Wang, W;Fäger, BM;Reisinger, H;Tokunaga, K;Cingöz, O;Sparrer, KMJ;Salker, MS;Hejnar, J;Kirchhoff, F;Trejbalova, K;Sauter, D; Inhibition of placental trophoblast fusion by guanylate-binding protein 5 Science advances 2025-05-09 [PMID: 40333975] (Bioassay, Human)

Bioinformatics

• Gene Symbol: PCSK7
• Uniprot:
  • Q16549()