Abstract
Proteoglycans are molecules that comprise a core protein to which at least one glycosaminoglycan (GAG) chain is attached, and a consideration of recombinant proteoglycan expression must operate under this simple definition. There are numerous systems currently available to express proteoglycans (and other complex glycoconjugates), but this chapter will focus on a system adopted by us to express proteoglycans and other extracellular matrix proteins in mammalian cells (1-5).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chan, D., Lamande, S. R., McQuillan, D. J., and Bateman, J. F. 1997. In vitro expression analysis of collagen biosynthesis and assembly. J. Biochem. Biophys. Meth. 36, 11–29.
Chan, D., Weng, Y. M., Hocking, A. M., Golub, S., McQuillan, D. J. and Bateman, J. F. 1996. Site-directed mutagenesis of human type X collagen. Expression of alpha1X) NC1, NC2, and helical mutations in vitro and in transfected cells. J. Biol. Chem. 271, 13,566–13,572.
Hocking, A. M., Strugnell, R., Ramamurthy, A. P., and McQuillan, D. J. 1996. Eukaryotic expression of recombinant biglycan. Post-translational processing and the importance of secondary structure for biological activity. J. Biol. Chem. 271, 19,571–19,577.
Krishnan, P., Hocking, A. M., Scholtz, J. M., Pace, C. N., Holik, K. K., and McQuillan, D. J. 1999. Distinct secondary structures of the leucine-rich repeat proteoglycans decorin and biglycan. Glycosylation-dependent conformational stability. J. Biol. Chem. 274, 10,945–10,950.
Ramamurthy, P., Hocking, A. M., and McQuillan, D. J. 1996. Recombinant decorin glycoforms. Purification and structure. J. Biol. Chem. 271, 19,578–19,584.
Hering, T. M., Kollar, J., Huynh, T. D., and Varelas, J. B. 1996. Purification and characterization of decorin core protein expressed in Escherichia coli as a maltose-binding protein fusion. Anal. Biochem. 240, 98–108.
Hildebrand, A., Romaris, M., Rasmussen, L. M., Heinegard, D., Twardzik, D. R., Border, W. A., and Ruoslahti, E. 1994. Interaction of the small interstitial proteoglycans biglycan, decorin and fibromodulin with transforming growth factor beta. Biochem. J. 302, 527–534.
Schonherr, E., Hausser, H., Beavan, L., and Kresse, H. 1995. Decorin-type I collagen interaction. Presence of separate core protein-binding domains. J. Biol. Chem. 270, 8877–8883.
Schonherr, E., Witsch-Prehm, P., Harrach, B., Robenek, H., Rauterberg, J., and Kresse, H. 1995. Interaction of biglycan with type I collagen. J. Biol. Chem. 270, 2776–2783.
Groffen, A. J., Buskens, C. A., Tryggvason, K., Veerkamp, J. H., Monnens, L. A., and van den Heuvel, L. P. 1996. Expression and characterization of human perlecan domains I and II synthesized by baculovirus-infected insect cells. Eur. J. Biochem. 241, 827–834.
Gu, J., Nakayama, Y., Nagai, K., and Wada, Y. 1997. The production and purification of functional decorin in a baculovirus system. Biochem. Biophys. Res. Commun. 232, 91–95.
Hochuli, E., Dobeli, H., and Schacher, A. 1987. New metal chelate adsorbent selective for proteins and peptides containing neighbouring histidine residues. J. Chromatogr. 411, 177–184.
Mackett, M., Smith, G. L., and Moss, B. 1985. The construction and characterization of vaccinia virus recombinants expressing foreign genes; in DNA Cloning, volume II (Glover, D. M., ed.), IRL, Oxford, UK, pp.191–212.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. M., Seidman, J. G., Smith, J. A., and Struhl, K., eds. 1987. Current Protocols in Molecular Biology. Wiley, New York, NY.
Elroy-Stein, O., Fuerst, T. R., and Moss, B. 1989. Cap-independent translation of mRNA conferred by encephalomyocarditis virus 5′ sequence improves the performance of the vaccinia virus/bacteriophage T7 hybrid expression system. Proc. Natl. Acad. Sci. (USA) 86, 6126–6130.
Fuerst, T. R., Niles, E. G., Studier, F. W., and Moss, B. 1986. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc. Natl. Acad. Sci. (USA) 83, 8122–8126.
Fuerst, T. R., Earl, P. L., and Moss, B. 1987. Use of a hybrid vaccinia virus-T7 RNA polymerase system for expression of target genes. Mol. Cell. Biol. 7, 2538–2544.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
McQuillan, D.J., Seo, NS., Hocking, A.M., McQuillan, C.I. (2001). Recombinant Expression of Proteoglycans in Mammalian Cells. In: Iozzo, R.V. (eds) Proteoglycan Protocols. Methods in Molecular Biology™, vol 171. Humana Press. https://doi.org/10.1385/1-59259-209-0:201
Download citation
DOI: https://doi.org/10.1385/1-59259-209-0:201
Publisher Name: Humana Press
Print ISBN: 978-0-89603-759-5
Online ISBN: 978-1-59259-209-8
eBook Packages: Springer Protocols