Abstract
The “a disintegrin and metalloproteinase with thrombospondin motifs” (ADAMTS) enzymes are secreted proteinases involved in development, blood clotting and the turnover of extracellular matrix. Manufacturing recombinant enzyme presents quite a challenge due to the presence of disulphide bridges, the large size and modular structure. A sub-group of these enzymes are known as “aggrecanases” and it is likely that they are involved in a number of pathologies related to increased turnover of the extracellular matrix, particularly in tissues where the concentration of proteoglycans is high, such as cartilage and the central nervous system. We have expressed three of these enzymes, ADAMTS-1, -4 and -5, in insect cells using plasmid-based systems.
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References
Huxley-Jones, J., Apte, S. S., Robertson, D. L., and Boot-Handford, R. P. (2005) The characterisation of six ADAMTS proteases in the basal chordate Ciona intestinalis provides new insights into the vertebrate ADAMTS family. Int J Biochem Cell Biol 37, 1838–1845.
Rawlings, N. D., Morton, F. R. and Barrett, A. J. (2006) MEROPS: the peptidase database. Nucleic Acids Res 34, D270–D272.
Jones, G. C. and Riley, G. P. (2005) ADAMTS proteinases: a multi-domain, multi-functional family with roles in extracellular matrix turnover and arthritis. Arthritis Res Ther 7, 160–169.
Flannery, C. R., Zeng, W., Corcoran, C., Collins-Racie, L. A., Chockalingam, P. S., Hebert, T., Mackie, S. A., McDonagh, T., Crawford, T. K., Tomkinson, K. N., LaVallie, E. R., and Morris, E. A. (2002) Autocatalytic cleavage of ADAMTS-4 (Aggrecanase-1) reveals multiple glycosaminoglycan-binding sites. J Biol Chem 277, 42775–42780.
Kuno, K. and Matsushima, K. (1998) ADAMTS-1 protein anchors at the extracellular matrix through the thrombospondin type I motifs and its spacing region. J Biol Chem 273, 13912–13917.
Vankemmelbeke, M. N., Jones, G. C., Fowles, C., Ilic, M. Z., Handley, C. J., Day, A. J., Knight, C. G., Mort, J. S., and Buttle, D. J. (2003) Selective inhibition of ADAMTS-1, -4 and -5 by catechin gallate esters. Eur J Biochem 270, 2394–2403.
Liu, C. J., Kong, W., Xu, K., Luan, Y., Ilalov, K., Sehgal, B., Yu, S., Howell, R. D., and Di Cesare, P. E. (2006) ADAMTS-12 associates with and degrades cartilage oligomeric matrix protein. J Biol Chem 281, 15800–15808.
Davis, T. R., Wickham, T. J., McKenna, K. A., Granados, R. R., Shuler, M. L., and Wood, H. A. (1993) Comparative recombinant protein production of eight insect cell lines. In Vitro Cell Dev Biol Anim 29A, 388–390.
Wickham, T. J., Davis, T., Granados, R. R., Shuler, M. L., and Wood, H. A. (1992) Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system. Biotechnol Prog 8, 391–396.
Wickham, T. J. and Nemerow, G. R. (1993) Optimization of growth methods and recombinant protein production in BTI-Tn-5B1-4 insect cells using the baculovirus expression system. Biotechnol Prog 9, 25–30.
Rhiel, M., Mitchell-Logean, C. M., and Murhammer, D. W. (1997) Comparison of Trichoplusia ni BTI-Tn-5B1-4 (high fivetrade mark) and Spodoptera frugiperda Sf-9 insect cell line metabolism in suspension cultures. Biotechnol Bioeng 55, 909–920.
Myers, E. W. and Miller, W. (1988) Optimal alignments in linear space. Comput Appl Biosci 4, 11–17.
Sambrook, J. and Russell, D. W. (2001). Purification of closed circular DNA by equilibrium centrifugation in CsCl-ethidium bromide gradients: continuous gradients. In Sambrook, J. and Russell, D. W. (eds.), Molecular cloning: a laboratory manual. New York: Cold Spring Harbour Laboratory Press, pp. 1.65–1.69.
Handley, C. J. and Buttle, D. J. (1995) Assay of proteoglycan degradation. Methods Enzymol 248, 47–58.
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Jones, G.C., Vankemmelbeke, M.N., Buttle, D.J. (2010). Expression of Recombinant ADAMTS in Insect Cells. In: Clark, I. (eds) Matrix Metalloproteinase Protocols. Methods in Molecular Biology, vol 622. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-299-5_5
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DOI: https://doi.org/10.1007/978-1-60327-299-5_5
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