Recombinant Human Marapsin/Pancreasin Protein, CF

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

Recombinant Human Marapsin/Pancreasin Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Gly-Arg-ThioBenzyl ester (Z-GR-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468. The specific activity is >2,000 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Marapsin/Pancreasin protein
  Ala23-Arg34, and Met35-Lys290 with a C-terminal 10-His tag
• Accession #: Q9BQR3
• N-terminal Sequence: Ala23 & Met35
• Structure / Form: Disulfide-linked heterodimer
• Protein/Peptide Type: Recombinant Enzymes
• Gene: PRSS27
• 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: 31 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: 39 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 reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile 25 mM MES and 100 mM NaCl, pH 6.5.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 250 mM NaCl, 0.05% (w/v) Brij-35, pH 8.0
  • Recombinant Human Marapsin/Pancreasin (rhMarapsin) (Catalog # 1988-SE)
  • Substrate: Z-Gly-Arg-SBzl (MP Biomedicals, Catalog # SB007)
  • 5,5’-Dithio-bis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130)
  • 96 well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhMarapsin to 0.4 ng/µL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer with 200 µM of DTNB.
  3. Load 50 µL of the diluted rhMarapsin into the plate, and start the reaction by adding 50 µL of the Substrate/DTNB mixture. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate mixture.
  4. Read in kinetic mode for 5 minutes at an absorbance of 405 nm.
  5. Calculate specific activity:

  Specific Activity (pmol/min/µg) = (Adjusted Vmax* (OD/min) x well volume (L) x 10^12 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:
  • rhMarapsin: 0.020 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 Marapsin/Pancreasin Protein, CF

• CAP-2
• CAPH2
• channel-activating protease 2
• EC 3.4.21
• EC 3.4.21.-
• EC 3.4.21.4
• EC 3.4.21.43
• Marapsin
• MPN
• MPNMarapsin
• Pancreasin
• protease, serine 27
• PRSS27
• serine protease 27

Background

Marapsin, pancreasin, and channel-activating protease 2 (CAP-2) are different names for the same serine protease that is expressed strongly in the pancreas (1, 2). The protein is synthesized with a signal peptide (amino acid residues 1-22), a pro peptide (residues 23-34) and a mature chain (residues 35-290) corresponding to the serine protease domain. The full-length protein was expressed and the secreted protein purified. The N-terminal sequencing results indicate that the purified protein is a disulfide bond-linked dimer formed between the pro peptide and the mature chain. The active enzyme has low activity against peptide substrates tested, but high activity against thioester substrates. The peptidase activity is inhibited by 20 mM benzamidine.

1. Bhagwandin, V.J. et al. (2003) J. Biol. Chem. 278:3363.
2. Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468.

Bioinformatics

• Gene Symbol: PRSS27
• Uniprot:
  • Q9BQR3()