Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF

R&D Systems | Catalog # 4007-ZN

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Mouse Meprin alpha Subunit/MEP1A Protein (4007-ZN)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

Sf 21 (baculovirus)

Accession Number

NP_032611

Structure / Form

Pro form

Applications

Enzyme Activity
View all Meprin alpha Subunit/MEP1A Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse Meprin alpha Subunit/MEP1A protein
Val34-Arg615, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Val34

Predicted Molecular Mass

68 kDa

SDS-PAGE

80 kDa, reducing conditions

Activity

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.

Formulation, Preparation, and Storage

4007-ZN
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: Meprin alpha Subunit/MEP1A

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).

References

  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.

Alternate Names

MEP1A, PPHA

Entrez Gene IDs

4224 (Human); 17287 (Mouse); 25684 (Rat)

Gene Symbol

MEP1A

UniProt

NP_032611

Additional Meprin alpha Subunit/MEP1A Products

Product Documents for Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Meprin alpha Subunit/MEP1A Protein, CF (4007-ZN):

Materials
  • 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

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