Recombinant Human THOP1 Protein, CF

R&D Systems | Catalog # 3439-ZN

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human THOP1 Protein (3439-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

E. coli

Accession Number

NP_003240

Applications

Enzyme Activity
View all Thimet Oligopeptidase/THOP1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Thimet Oligopeptidase/THOP1 protein
Lys2-Cys689, with an N-terminal Met and 6-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

Met

Predicted Molecular Mass

80 kDa

SDS-PAGE

80 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic peptide substrate, (7-methoxycoumarin-4-yl)acetyl-Pro-Leu-Gly-Pro-D-Lys(2,4-dinitrophenyl)-OH or Mca-PLGPK(Dnp)-OH.
The specific activity is >250 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

3439-ZN
Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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: Thimet Oligopeptidase/THOP1

Thimet Oligopeptidase/THOP1, also known as endopeptidase EC 3.4.24.15 (EP24.15), is a zinc peptidase of the M3 family that also includes neurolysin/EC 3.4.24.16 and mitochondrial intermediate peptidase (1). Widely expressed by mammalian tissues and reported to present in different subcellular locations, THOP1 is primarily a cytoplasmic enzyme. It is capable of hydrolyzing a number of bioactive peptides and peptides released by the proteosome, limiting antigenic presentation by MHC class I molecules (1-3). THOP1 also interacts with angiotensin II type I receptor and bradykinin B2 receptor (4). The optimal activity of the purified THOP1 may or may not require the presence of a reducing agent, depending upon the source of the enzyme and the purification method used.

References

  1. Barrett, A.J. and J.-M. Chen (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. eds. p. 352, Elsevier Academic Press, San Diego.
  2. Ray, K. et al. (2004) J. Biol. Chem. 279:20480.
  3. Saric, T. et al. (2004) J. Biol. Chem. 279:46723.
  4. Shivakumar, B.R. et al. (2005) Cell Biochem. Funct. 23:195.

Alternate Names

EP24.15

Entrez Gene IDs

7064 (Human); 50492 (Mouse); 64517 (Rat)

Gene Symbol

THOP1

UniProt

NP_003240

Additional Thimet Oligopeptidase/THOP1 Products

Product Documents for Recombinant Human THOP1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human THOP1 Protein, CF

For research use only

Related Research Areas

Metalloproteases and Regulators

Citations for Recombinant Human THOP1 Protein, CF

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Protocols

View specific protocols for Recombinant Human THOP1 Protein, CF (3439-ZN):

Materials
  • Assay Buffer: 25 mM Tris, 150 mM NaCl, pH 7.5
  • Recombinant Human Thimet Oligopeptidase/THOP1 (rhTHOP1) (Catalog # 3439-ZN)
  • Substrate: MCA-Pro-Leu-Gly-Pro-D-Lys-(DNP)-OH (Bachem, Catalog # M-2270); 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Spectramax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhTHOP1 to 0.5 ng/µL in Assay Buffer.
  2. Dilute Substrate to 20 µM in Assay Buffer.
  3. Load 50 µL of the 0.5 ng/µL rhTHOP1 in a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
  4. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. 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:
  • rhTHOP1: 0.025 µg
  • Substrate: 10 µM

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