ADAMTS4 (a disintegrin and metalloproteinase with thrombospondin motifs 4), also known as aggrecanase-1, is a member of the family of secreted zinc proteases with a multi-domain structure (1-3). The protein precursors consist of a signal peptide and the following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine-rich, and spacer. It is the only ADAMTS identified that has one TS type I motif. It is an active protease effectively cleaving alpha -2-macroglobulin and aggrecan at multiple sites, and is inhibited by TIMP-3 with inhibition constants in subnanomolar range (4-6). It receives great attention due to the elevation in its mRNA level after treatment with Interleukin-1 (7). However, in a mouse model of osteoarthritis, ADAMTS4 knock-out mice did not exhibit any significant protective effect (8). The purified rhADAMTS4 starts at the catalytic domain and ends before the spacer domain. If desired, the aggrecanase activity can be inhibited by 5 mM 1 10‑phenanthroline and Recombinant Human TIMP‑3 (Catalog # http://www.rndsystems.com/product_results.aspx?k=973-TM">973-TM).
Recombinant Human ADAMTS4 Protein, CF
R&D Systems | Catalog # 4307-AD
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Key Product Details
- R&D Systems CHO-derived Recombinant Human ADAMTS4 Protein (4307-AD)
- 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
CHO
Accession Number
Structure / Form
Recombinant Human ADAMTS4 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
Applications
Enzyme Activity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human ADAMTS4 protein
Phe213-Cys685, with a C-terminal 10-His tag
Phe213-Cys685, with a C-terminal 10-His tag
Purity
>90%, 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
Phe213
Predicted Molecular Mass
53 kDa
SDS-PAGE
58 kDa, reducing conditions
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Abz-TEGEARGSVI-Dap(Dnp)-KK-NH2.
The specific activity is >7 pmol/min/μg, as measured under the described conditions.
The specific activity is >7 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
4307-AD
| Formulation | Supplied as a 0.2 μm filtered solution in Sodium Acetate, CaCl2 and NaCl. |
| 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.
|
Background: ADAMTS4
References
- Totorella, M. D. et al. (1999) Science 284:1664.
- Porter, S. et al. (2005) Biochem. J. 386:15.
- Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
- Tortorella, M. D. et al. (2004) J. Biol. Chem. 279:17554.
- Struglics, A. et al. (2006) Osteoarthritis Cartilage. 14:101.
- Kashiwagi, M. et al. (2001) J. Biol. Chem. 276:12501.
- Pratta, M. A. et al. (2003) Arthritis Reum. 48:119.
- Glasson, S. S. et al. (2004) Arthritis Reum. 50:2547.
Long Name
A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 4
Alternate Names
ADMP-1, Aggrecanase 1
Gene Symbol
ADAMTS4
UniProt
Additional ADAMTS4 Products
Product Documents for Recombinant Human ADAMTS4 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human ADAMTS4 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ADAMTS4 Protein, CF
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Protocols
View specific protocols for Recombinant Human ADAMTS4 Protein, CF (4307-AD):
Materials
- Assay Buffer: 50 mM HEPES, 50 mM NaCl, 1 mM CaCl2, 0.05% Brij-35, pH 7.5
- Recombinant Human ADAMTS4 (Catalog # 4307-AD)
- Substrate: WAAG-3R (Anaspec, Catalog # 60431-1), 2 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhADAMTS4 to 10 µg/mL in Assay Buffer.
- Dilute Substrate to 50 µM in Assay Buffer.
- Load 50 µL of 10 µg/mL of rhADAMTS4 into a plate, and start the reaction by adding 50 µL of 50 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
- Read at excitation and emission wavelengths of 340 nm and 420 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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 Abz-Gly-OH (Bachem, Catalog # E-2920).
- rhADAMTS4: 0.5 µg
- Substrate: 25 µM
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