Recombinant Human ADAMTS1 Protein, CF

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

Recombinant Human ADAMTS1 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave Recombinant Human Aggrecan G1-IGD-G2 Domains (Catalog # 1220-PG).
• Source: Mouse myeloma cell line, NS0-derived human ADAMTS1 protein
  Phe253-Ala734, with a C-terminal 10-His tag
• Accession #: AAD48080
• N-terminal Sequence: Phe253
• Structure / Form: Recombinant Human ADAMTS1 is prone to proteolytic cleavage at C-terminus. The predominant form of the purified protein lacks the His tag.
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ADAMTS1
• 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

• Theoretical MW: 54 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: 66 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 Sodium Acetate, CaCl₂ and NaCl.
• 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 Human Aggrecan G1-IGD-G2 (rhAggrecan) Domains (Catalog # 1220-PG)
  • Recombinant Human ADAMTS1 (rhADAMTS1) (Catalog # 2197-AD)
  • SDS-PAGE with Silver Staining
  1. Dilute rhAggrecan to 200 µg/mL in Assay Buffer.
  2. Dilute rhADAMTS1 to 56 µg/mL in Assay Buffer.
  3. Mix 25 µL of 200 µg/mL rhAggrecan, 10 µL of 56 µg/mL rhADAMTS1, and 15 µL of Assay Buffer.
  4. Incubate at 37 °C for 24 hours.
  5. Include controls containing equal volumes of 200 µg/mL rhAggrecan and Assay Buffer. Incubate one tube at 37 °C and the other at -20 °C for 24 hours.
  6. Stop the reaction by mixing equal volumes of the incubated samples and reducing gel loading buffer.
  7. Analyze the cleavage by SDS PAGE followed by protein silver staining [or Western blot using anti-human aggrecan antibodies (Catalog # AF1220 and MAB1220)].
  Per Reaction:
  • rhADAMTS1: 11.2 µg/mL
  • rhAggrecan: 100 µg/mL

Notes

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

Alternate Names for Recombinant Human ADAMTS1 Protein, CF

• a disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif, 1
• ADAM metallopeptidase with thrombospondin type 1 motif, 1
• ADAM-TS 1
• ADAMTS1
• ADAM-TS1
• ADAMTS-1
• C3-C5
• METH1
• METH-1

Background

ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1), also known as METH1, is the founding member of the family of secreted zinc proteases with a multi-domain structure (1-3). The protein precursors consist of signal peptide and following domains: pro, catalytic, disintegrin-like, TS type 1 motif, cysteine‑rich, spacer and a variable number of TS type 1 motifs. Based on their substrate specificity, ADAMTS1 and associated family members may be key enzymes in degradation of cartilage leading to inflammation and arthritis (4). It is an active protease cleaving alpha -2-macroglobulin (5), aggrecan (6), and versican (7). Compared to ADAMTS4 (aggrecanase 1) and ADAMTS5 (aggrecanase 2), the aggrecanase activity of ADAMTS1 is lower. However, its activity can be enhanced by the binding of cofactor such as fibulin-1 (8). The aggrecanase activity can be inhibited using 5 mM 1,10 Phenanthroline. ADAMTS1 is essential for normal growth and the structure and function of the kidneys, adrenal glands and female reproductive organs (9). It also plays an important role in atherosclerosis (10). It has been shown to inhibit endothelial cell proliferation by direct binding and sequestration of VEGF165 and to inhibit fibroblast migration at high concentrations by binding to FGF-2 (11, 12). The purified recombinant human ADAMTS1 starts at the N‑terminus of the catalytic domain and ends in the beginning of the spacer region.

1. Vazquez, F. et al. (1999) J. Biol. Chem. 274:23349.
2. Kuno, K. et al. (1997) J. Biol. Chem. 272:556.
3. Porter, S. et al. (2005) Biochem. J. 386:15.
4. Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
5. Kuno, K. et al. (1999) J. Biol. Chem. 274:18821.
6. Kuno, K. et al. (2000) FEBS Lett. 478:241.
7. Russel, D. L. et al. (2003) J. Biol. Chem. 278:42330.
8. Lee, N., et al. (2005) J. Biol. Chem. 280:34796.
9. Shindo, T., et al. (2000) J. Clin. Invest. 105:1345.
10. Wight, T.N. (2005) Arterioscler Thromb. Vasc. Biol. 25:12.
11. Luque, A. et al. (2003) J. Biol. Chem. 278:23656.
12. Krampert, M. et al. (2005) J. Biol. Chem. 280:23844.

Publications for ADAMTS1 (2197-AD)(4)

Publications using 2197-AD

• T Fontanil, S ?lvarez-Te, M? Villaronga, Y Mohamedi, L Solares, A Moncada-Pa, JA Vega, O Garc¡a-Su , M P‚rez-Bast, JM Garc¡a-Ped, AJ Obaya, S Cal Cleavage of Fibulin-2 by the aggrecanases ADAMTS-4 and ADAMTS-5 contributes to the tumorigenic potential of breast cancer cells Oncotarget, 2017;0(0):. 2017 [PMID: 28099917] (Enzyme Assay, Human)
• Lemarchant S, Pomeshchik Y, Kidin I, Karkkainen V, Valonen P, Lehtonen S, Goldsteins G, Malm T, Kanninen K, Koistinaho J ADAMTS-4 promotes neurodegeneration in a mouse model of amyotrophic lateral sclerosis. Mol Neurodegener, 2016;11(1):10. 2016 [PMID: 26809777] (Bioassay, Mouse)
• Wang Z, Luo J, Iwamoto S, Chen Q Matrilin-2 is proteolytically cleaved by ADAMTS-4 and ADAMTS-5. Molecules, 2014;19(6):8472-87. 2014 [PMID: 24959676] (Bioassay, Primate - Chlorocebus aethiops (African Green Monkey))
• Rocks N, Paulissen G, Quesada-Calvo F, Munaut C, Gonzalez ML, Gueders M, Hacha J, Gilles C, Foidart JM, Noel A, Cataldo DD ADAMTS-1 metalloproteinase promotes tumor development through the induction of a stromal reaction in vivo. Cancer Res., 2008;68(22):9541-50. 2008 [PMID: 19010931] (Bioassay, Human)

Bioinformatics

• Gene Symbol: ADAMTS1
• Uniprot:
  • AAD48080()