Recombinant Human PCPE-1 Protein, CF

R&D Systems | Catalog # 2627-PE

R&D Systems
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Key Product Details

  • R&D Systems NS0-derived Recombinant Human PCPE-1 Protein (2627-PE)
  • 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

NS0

Accession Number

Q15113

Applications

Enzyme Activity
View all PCPE-1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human PCPE-1 protein
Met1-Asp449, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

No result, Gln26 expected

Predicted Molecular Mass

47 kDa

SDS-PAGE

51-57 kDa, reducing conditions

Activity

Measured by the stimulation of procollagen type I processing by BMP-1.
>50% of the full-length procollagen, in the presence of Recombinant Human PCPE-1, is processed by BMP-1, as measured under the described conditions.

Formulation, Preparation, and Storage

2627-PE
Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: PCPE-1

Procollagen C‑endopeptidase enhancers, known as PCPEs or PCOLCEs, are secreted extracellular matrix glycoproteins that consist of two CUB domains and one NTR domain. They are known to stimulate enzymatic cleavage of procollagens I‑III by the BMP‑1/tolloid family of metalloproteases, also known as procollagen C‑proteinases (1). PCPE‑1 is expressed primarily by interstitial connective tissues such as tendons, calvaria, and skin (2). Although BMP‑1/tolloid proteinases are involved in processing of multiple extracellular proteins, the enhancer activity of PCPE‑1 is specific to procollagens since it has no effect on BMP‑1/tolloid cleavage of other substrates (3). It is thought that PCPE‑1 enhances cleavage of procollagens by binding to the substrate and inducing a conformation change in the substrate (3), although interaction between PCPE‑1 and full‑length BMP‑1/tolloid proteinases has also been reported (4).

References

  1. Steiglitz, B.M. et al. (2002) J. Biol. Chem. 277:49820.
  2. Kessler, E. et al. (1990) Biochem. Biophys. Res. Commun. 173:81.
  3. Moali, C. et al. (2005) J. Biol. Chem. 280:24188.
  4. Ge, G. et al. (2006) J. Biol. Chem. 281:10786.

Long Name

Procollagen C-Proteinase Enhancer-1

Alternate Names

PCOLCE, PCPE1

Entrez Gene IDs

5118 (Human); 18542 (Mouse)

Gene Symbol

PCOLCE

UniProt

Q15113

Additional PCPE-1 Products

Product Documents for Recombinant Human PCPE-1 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 PCPE-1 Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Related Research Areas

Metalloproteases and Regulators

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Protocols

View specific protocols for Recombinant Human PCPE-1 Protein, CF (2627-PE):

Materials
  • Dilution Buffer: 50 mM Tris, 5 mM CaCl2, pH 7.5
  • Assay Buffer: 50 mM Tris, 5 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
  • Recombinant Human PCPE-1 (rhPCPE-1) (Catalog # 2627-PE)
  • Recombinant Human Pro Collagen I  alpha 1 (rhCOL1A1) (Catalog # 6220-CL)
  • Recombinant Human BMP‑1/PCP (rhBMP-1) (Catalog # 1927-ZN)
  • Reducing SDS-PAGE Sample Buffer (3X or greater)
  • SDS-PAGE or Western Blot
  1. Dilute rhCOL1A1 to 75 µg/mL in Dilution Buffer.
  2. Dilute rhPCPE-1 to 15 µg/mL in Assay Buffer.
  3. In a tube, combine equal volumes (a minimum of 15 µL of each is required for this assay) of diluted rhCOL1A1 and diluted rhPCPE‑1. Prepare samples in duplicate (one replicate to be used as a control).
  4. Incubate vials at 37 °C for 30 minutes.
  5. Dilute rhBMP-1 to 1.875 µg/mL in Assay Buffer.
  6. Add one equivalent (1/2 the total volume of rhCOL1A1/rhPCPE-1 mixtures) of diluted rhBMP-1 to each reaction tube. Prevent the reaction from occurring in the control duplicates by immediately mixing with reducing gel buffer (see step 8 for volumes).
  7. Incubate reaction tubes at 37 °C for 30 minutes.
  8. After incubation, combine 40 µL of each reaction mixture and control with 20 µL of reducing SDS-PAGE gel buffer. Mix and incubate samples at 95-100 °C for 3-5 minutes.
  9. Load 30 µL per lane and analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
  10. Activity calculation:
     % Cleavage = [1 -  % full-length rhCOL1A1 (reaction) ] x 100%
% full-length rhCOL1A1 (control)

Per Lane:

  • rhPCPE-1: 0.1 µg
  • rhCOL1A1: 0.5 µg
  • rhBMP-1: 0.0125 µg

FAQs

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