Combine 25 µL of 200 µg/mL rhAggrecan with 25 µL of 20 µg/mL rmADAMTS1 in a reaction tube.
Incubate the reaction mixture for 24 hours at 37 °C.
For Controls: Combine 25 µL of rhAggrecan solution and 25 µL of Assay Buffer into two tubes. Incubate one tube at 37 °C and the other at -20 °C for 24 hours.
Stop the reaction by adding reduced SDS-PAGE sample buffer.
Analyze the cleavage by SDS PAGE followed by protein staining and/or Western blot.
rmADAMTS1: 10 µg/mL (0.5 µg)
rhAggrecan: 100 µg/mL (5 µg)
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 multidomain structure (1-3). The protein precursor consists of a signal peptide and the following domains: pro, catalytic, disintegrinlike, TS type 1 motif, cysteine rich, spacer and a variable number of thrombospondin type 1 motifs. Based on their substrate specificity, ADAMTS1 and associated family members may be key enzymes in the degradation of cartilage leading to inflammation and arthritis (4). It is an active protease cleaving alpha 2macroglobulin (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 a cofactor such as fibulin1 (8). 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 rmADAMTS1 starts at the N terminus of the catalytic domain and ends in the Cys-rich domain.
Vazquez, F. et al. (1999) J. Biol. Chem. 274:23349.
Kuno, K. et al. (1997) J. Biol. Chem. 272:556.
Porter, S. et al. (2005) Biochem. J. 386:15.
Nagase, H. and M. Kashiwagi (2003) Arthritis Res. Ther. 5:94.
Kuno, K. et al. (1999) J. Biol. Chem. 274:18821.
Kuno, K. et al. (2000) FEBS Lett. 478:241.
Russel, D. L. et al. (2003) J. Biol. Chem. 278:42330.
Lee, N. et al. (2005) J. Biol. Chem. 280:34796.
Shindo, T. et al. (2000) J. Clin. Invest. 105:1345.
Krampert, M. et al. (2005) J. Biol. Chem. 280:23844.
A Disintegrin-like and Metalloproteinase Domain with Thrombospondin Motifs 1
Entrez Gene IDs:
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
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The data collected includes not only links to publications in PubMed,
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