Recombinant Mouse MMP-7 Protein, CF

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

Recombinant Mouse MMP-7 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH₂ (Catalog # ES010). The specific activity is >2000 pmol/min/µg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived mouse MMP-7 protein
  Met1-Leu264
• Accession #: AAA99983
• N-terminal Sequence: Leu18
• Structure / Form: Pro form
• Protein/Peptide Type: Recombinant Enzymes
• Gene: Mmp7
• Purity: >90%, 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: 28 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: 31 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 MES, NaCl, CaCl₂ and Glycerol.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Mouse MMP-7 (rmMMP-7) (Catalog # 2967-MP)
  • p-aminophenylmercuric acetate (APMA) (Sigma, Catalog # A-9563), 100 mM stock in DMSO
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂, (Catalog # ES010)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmMMP-7 to 100 μg/mL in Assay Buffer.
  2. Activate rmMMP-7 by adding APMA to a final concentration of 1 mM.
  3. Incubate at 37 °C for 1 hour.
  4. Dilute activated rmMMP-7 to 0.4 ng/μL in Assay Buffer.
  5. Dilute Substrate to 120 μM in Assay Buffer.
  6. In a plate, load 50 μL of 0.4 ng/μL rmMMP-7 and start the reaction by adding 50 μL of 120 μM Substrate. Include a Substrate Blank containing 50 μL Assay Buffer and 50 μL of 120 μM Substrate.
  7. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  8. 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:
  • rmMMP-7: 0.020 μg
  • Substrate: 60 μM

Notes

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

Alternate Names for Recombinant Mouse MMP-7 Protein, CF

• EC 3.4.24
• EC 3.4.24.23
• Matrilysin
• matrin
• matrix metallopeptidase 7 (matrilysin, uterine)
• matrix metalloproteinase 7 (matrilysin, uterine)
• Matrix metalloproteinase-7
• MMP7
• MMP-7
• MPSL1
• Pump-1 protease
• PUMP1
• PUMP-1
• uterine matrilysin
• Uterine metalloproteinase

Background

Matrix metalloproteinases (MMPs) are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade all the components of the extracellular matrix. MMP-7 (matrilysin) is expressed in epithelial cells of normal and diseased tissues, and is capable of digesting a large series of proteins of the extracellular matrix including collagen IV and X, gelatin, casein, laminin, aggrecan, entactin, elastin and versican (1). MMP-7 is implicated in the activation of other proteinases such as plasminogen, MMP-1, MMP-2, and MMP-9. In addition to its roles in connective tissue remodeling and cancer, MMP-7 also regulates intestinal alpha ‑defensin activation in innate host defense, releases tumor necrosis factor-alpha in a model of herniated disc resorption, and cleaves FasL to generate a soluble form in a model of prostate involution. Structurally, MMP-7 is the smallest of the MMPs and consists of two domains: a pro-domain that is cleaved upon activation and a catalytic domain containing the zinc-binding site.

1. Woessner, J.F. (2004) in Handbook of Proteolytic Enzymes, Barrett, A.J. et al. eds. p. 532.

Publications for MMP-7 (2967-MP)(3)

Publications using 2967-MP

• Levengood, MR;Carosino, CM;Zhang, X;Lucas, S;Ortiz, DJ;Westendorf, L;Chin, AP;Martin, AD;Wong, A;Hengel, SM;Sun, H;Zeng, W;Yumul, R;Dominguez, MM;Chen, Y;Zheng, JH;Karlsson, CAB;Trang, VH;Senter, PD;Gardai, SJ; Preclinical development of SGN-CD47M: Protease-activated antibody technology enables selective tumor targeting of the innate immune checkpoint receptor CD47 Molecular cancer therapeutics 2024-10-28 [PMID: 39463068] (Bioassay, N/A)
• Kon S, Nakayama Y, Matsumoto N, Ito K, Kanayama M, Kimura C, Kouro H, Ashitomi D, Matsuda T, Uede T A novel cryptic binding motif, LRSKSRSFQVSDEQY, in the C-terminal fragment of MMP-3/7-cleaved osteopontin as a novel ligand for alpha9beta1 integrin is involved in the anti-type II collagen antibody-induced arthritis. PLoS ONE, 2014-12-29;9(12):e116210. 2014-12-29 [PMID: 25545242] (Bioassay, Mouse)
• Suryawanshi A, Mulik S, Sharma S Ocular neovascularization caused by herpes simplex virus type 1 infection results from breakdown of binding between vascular endothelial growth factor A and its soluble receptor. J. Immunol., 2011-02-16;186(6):3653-65. 2011-02-16 [PMID: 21325621] (Enzyme Assay, Mouse)

Bioinformatics

• Gene Symbol: Mmp7
• Uniprot:
  • AAA99983()