Recombinant Human ADAMTS5 Protein, CF

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

Recombinant Human ADAMTS5 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave the fluorogenic peptide substrate, Abz-TEGEARGSVI-Dap(Dnp)-KK-NH2. The specific activity is >10 pmol/min/μg, as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human ADAMTS5 protein
  Ser262-Pro622, with a C-terminal 10-His tag
• Accession #: Q9UNA0
• N-terminal Sequence: Ser262
• Structure / Form: Recombinant Human ADAMTS5 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: ADAMTS5
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 41 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: 51 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, -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in Sodium Acetate, CaCl₂ and NaCl.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 200 μg/mL in sterile, deionized water.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 100 mM NaCl, 5 mM CaCl₂, 0.05% Brij-35, pH 7.5
  • Recombinant Human ADAMTS5 (rhADAMTS5) (Catalog # 2198-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
  1. Dilute rhADAMTS5 to 10 µg/mL in Assay Buffer.
  2. Dilute Substrate to 50 µM in Assay Buffer.
  3. Load 50 µL of 10 µg/mL of rhADAMTS5 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.
  4. Read at excitation and emission wavelengths of 340 nm and 420 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 Abz-Gly-OH (Bachem, Catalog # E-2920).

  Per Well:
  • rhADAMTS5: 0.5 µg
  • Substrate: 25 µM

Notes

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

Alternate Names for Recombinant Human ADAMTS5 Protein, CF

• A disintegrin and metalloproteinase with thrombospondin motifs 11
• a disintegrin-like and metalloprotease (reprolysin type) with thrombospondintype 1 motif, 5 (aggrecanase-2)
• ADAM metallopeptidase with thrombospondin type 1 motif, 5
• ADAM-TS 11
• ADAM-TS 5
• ADAMTS11
• ADAMTS-11
• ADAMTS5
• ADAM-TS5
• ADAMTS-5
• ADMP2
• ADMP-2
• ADMP-2A disintegrin and metalloproteinase with thrombospondin motifs 5
• Aggrecanase 2
• aggrecanase-2
• disintegrin-like and metalloprotease with thrombospondin type 1 motif, 510ADAMTS11FLJ36738
• EC 3.4.24
• EC 3.4.24.-
• EC 3.4.24.14
• EC 3.4.24.82

Background

ADAMTS5 (a disintegrin and metalloproteinase with thrombospondin motifs 5), also known as aggrecanase-2 and ADAMTS11, is a member of the family of secreted zinc proteases with a multi-domain structure (1, 2). 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. ADAMTS5 is an active protease effectively cleaving alpha 2-macroglobulin (3), aggrecan (4), and brevican (5), and is inhibited by TIMP-3 with inhibition constants in the subnanomolar range (6). Based on the murine model studies (7, 8), this protease may be a key enzyme in the degradation of cartilage leading to osteoarthritis and rheumatoid arthritis. The purified rhADAMTS5 starts at the N-terminus of the catalytic domain and ends at the C-terminus of the TSP-1 domain. The amino acid sequence of rhADAMTS5 is 98%, 97%, and 96% identical to that of canine, bovine, and mouse/rat. The aggrecanase activity can be inhibited by 5 mM 1,10-phenanthroline and recombinant human TIMP-3 (Catalog # 973-TM).

1. Porter, S. et al. (2005) Biochem. J. 386:15.
2. Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
3. Tortorella, M.D. et al. (2004) J. Biol. Chem. 279:17554.
4. Vankemmelbeke, M.N. et al. (2001) Eur. J. Biochem. 268:1259.
5. Nakada, M. et al. (2005) Acta Neurophathol (Berl). 110:239.
6. Kashiwagi, M. et al. (2001) J. Biol. Chem. 276:12501.
7. Glasson, S.S. et al. (2005) Nature, 434: 644.
8. Stanton, H. et al. (2005) Nature, 434:648.

Publications for ADAMTS5 (2198-AD)(11)

Publications using 2198-AD

• Kuczynski-Noyau, L;Karmann, S;Alberton, P;Martinez-Corral, I;Nampoothiri, S;Sauvé, F;Lhomme, T;Quarta, C;Apte, SS;Bouret, S;Aszodi, A;Rasika, S;Ciofi, P;Dam, J;Prévot, V;Mattot, V; A plastic aggrecan barrier modulated by peripheral energy state gates metabolic signal access to arcuate neurons Nature communications 2024-08-07 [PMID: 39112471] (In vivo assay, Mouse)
• DC Kirman, B Renganatha, WK Chui, MW Chen, NA Kaya, R Ge Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis Cell Death & Disease, 2022-02-23;13(2):172. 2022-02-23 [PMID: 35197459] (Bioassay, Human)
• S Taylor, M Whitfield, J Barratt, A Didangelos The Metalloproteinase ADAMTS5 Is Expressed by Interstitial Inflammatory Cells in IgA Nephropathy and Is Proteolytically Active on the Kidney Matrix J. Immunol., 2020-09-11;0(0):. 2020-09-11 [PMID: 32917786] (Bioassay, Rat)
• T Fontanil, Y Mohamedi, A Moncada-Pa, T Cobo, JA Vega, JL Cobo, O García-Suá, J Cobo, ÁJ Obaya, S Cal Neurocan is a New Substrate for the ADAMTS12 Metalloprotease: Potential Implications in Neuropathies Cell. Physiol. Biochem., 2019-01-01;52(5):1003-1016. 2019-01-01 [PMID: 30977985] (Bioassay, Mouse)
• F Seuffert, D Weidner, W Baum, G Schett, M Stock Upper zone of growth plate and cartilage matrix associated protein protects cartilage during inflammatory arthritis Arthritis Res. Ther., 2018-05-02;20(1):88. 2018-05-02 [PMID: 29720262] (Bioassay, Human)
• F Li, R Song, L Ao, TB Reece, JC Cleveland, N Dong, DA Fullerton, X Meng ADAMTS5 Deficiency in Calcified Aortic Valves Is Associated With Elevated Pro-Osteogenic Activity in Valvular Interstitial Cells Arterioscler. Thromb. Vasc. Biol., 2017-05-25;0(0):. 2017-05-25 [PMID: 28546218] (Bioassay, Human)
• 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, 2017-02-21;0(0):. 2017-02-21 [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, 2016-01-25;11(1):10. 2016-01-25 [PMID: 26809777] (Bioassay, Mouse)
• He Y, Zheng Q, Jiang M, Sun S, Christiansen T, Kassem M, Karsdal M, Bay-Jensen A The effect of protease inhibitors on the induction of osteoarthritis-related biomarkers in bovine full-depth cartilage explants. PLoS ONE, 2015-04-24;10(0):. 2015-04-24 [PMID: 25909781] (Western Blot)
• Kumar S, Sharghi-Namini S, Rao N, Ge R ADAMTS5 functions as an anti-angiogenic and anti-tumorigenic protein independent of its proteoglycanase activity. Am J Pathol, 2012-07-13;181(3):1056-68. 2012-07-13 [PMID: 22796434] (Bioassay, Human)
• Devel L, Beau F, Amoura M, Vera L, Cassar-Lajeunesse E, Garcia S, Czarny B, Stura E, Dive V Simple pseudo-dipeptides with a P2&#039; glutamate: a novel inhibitor family of matrix metalloproteases and other metzincins. J Biol Chem, 2012-06-11;287(32):26647-56. 2012-06-11 [PMID: 22689580] (Enzyme Assay)

Bioinformatics

• Gene Symbol: ADAMTS5
• Uniprot:
  • Q9UNA0()