Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF

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

Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH (Catalog # ES007). The specific activity is >400 pmol/min/µg, as measured under the described conditions.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Meprin alpha Subunit/MEP1A protein
  Val34-Arg615, with a C-terminal 10-His tag
• Accession #: NP_032611
• N-terminal Sequence: Val34
• Structure / Form: Pro form
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Mep1a
• 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: 68 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: 80 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 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, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Assay Buffer: 50 mM Tris, 1 M NaCl, pH 9.0
  • Recombinant Mouse Meprin alpha Subunit/MEP1A (rmMEP1A) (Catalog # 4007-ZN)
  • Trypsin (Sigma, Catalog # T-1426)
  • AEBSF (Catalog # EI001), 100 mM stock in deionized water
  • Substrate: MCA-Tyr-Val-Ala-Asp-Ala-Pro-Lys(DNP)-OH (Catalog # ES007)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmMEP1A to 100 µg/mL in Activation Buffer with Trypsin at 0.1 µg/mL.
  2. Incubate for 45 minutes at 37 °C.
  3. Stop Trypsin activation by adding an equal volume of AEBSF at 2 mM in Activation Buffer and mixing well.
     Dilute Substrate to 40 µM in Assay Buffer.
  4. Dilute activated rmMEP1A to 0.8 µg/mL in Assay Buffer.
  5. Load in a plate 50 µL of 0.8 µg/mL rmMEP1A, and start the reaction by adding 50 µL of 40 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 40 µM Substrate.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

  Per Well:
  • rmMEP1A: 0.04 µg
  • Substrate: 20 µM

Notes

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

Alternate Names for Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF

• bA268F1.1 (meprin A alpha (PABA peptide hydrolase))
• EC 3.4.24
• EC 3.4.24.18
• endopeptidase-2
• MEP1A
• meprin A subunit alpha
• meprin A, alpha (PABA peptide hydrolase)
• Meprin alpha Subunit
• N-benzoyl-L-tyrosyl-P-amino-benzoic acid hydrolase subunit alpha
• PABA peptide hydrolase
• PPH alpha
• PPHA

Background

Meprins are multimeric proteases composed of alpha and beta subunits, which are members of the astacin family of zinc endopeptidases (1, 2). Both subunits form disulfide‑linked homo‑ or hetero‑oligomers, which are also referred to as Meprin A (composed of alpha subunits with or without beta subunits) and Meprin B (composed of beta subunits only) (3). Although the two subunits share 42% identity in their amino acid sequence, they differ significantly in their oligomeric structure, post‑translational processing and subsequently cellular location, and substrate and peptide bond specificity (4). The 760 amino acid sequence of mouse meprin alpha subunit precursor consists of a signal peptide (residues 1‑33), a pro region (residues 34‑77), and a mature chain (residues 78‑760) containing the following domains, catalytic (residues 78‑275), MAM (residues 276‑445), MATH (residues 447‑607), EGF‑like (residues 684‑724), transmembrane (residues 727‑754), and cytoplasmic (residues 755‑760) (5). The pro enzyme terminating at residue 615 was expressed and the secreted protein purified from conditioned medium. The molecular masses of recombinant mouse MEP1A are similar to those observed for the alpha subunit of rat Meprin A (6).

1. Bond, J.S. and R.J. Beynon (1995) Protein Sci. 4:1247.
2. Stocker, W. et al. (1995) Protein Sci. 4:823.
3. Bertenshaw, G.P. et al. (2001) J. Biol. Chem. 276:13248.
4. Ishmael, F.T. et al. (2005) J. Biol. Chem. 280:13895.
5. Jiang, W. et al. (1992) J. Biol. Chem. 267:9185.
6. Bertenshaw, G.P. et al. (2003) J. Biol. Chem. 278:2522.

Publications for Meprin alpha Subunit/MEP1A (4007-ZN)(2)

Publications using 4007-ZN

• Wang, CS;Zhong, Q;Wang, ST;Martin-Alonso, C;Neaher, S;Patel, S;Parisi, T;Kirkpatrick, JD;Sequist, LV;Jacks, TE;Bhatia, SN; Multimodal profiling of proinflammatory protease activity identifies caspase-1 as a target for lung cancer interception bioRxiv : the preprint server for biology 2025-07-02 [PMID: 40631193] (FRET, Mouse)
• Pankow K, Wang Y, Gembardt F, Krause E, Sun X, Krause G, Schultheiss HP, Siems WE, Walther T Successive action of meprin A and neprilysin catabolizes B-type natriuretic peptide. Circ. Res., 2007-09-06;101(9):875-82. 2007-09-06 [PMID: 17823376] (Bioassay)

Bioinformatics

• Gene Symbol: Mep1a
• Uniprot:
  • NP_032611()