title: Eukaryotic expression and purification of recombinant extracellular matrix proteins carrying the strep II tag.
abstract: For recombinant expression of extracellular matrix (ECM) proteins or their individual domains, the use of transformed mammalian cells offers two major advantages. First, eukaryotic expression can be expected under optimum conditions to produce a large proportion of correctly folded molecules. ECM proteins are made from a group of 25 structurally known (Rev. Biophys. 29:119-167, 1996) and about 200 cDNA derived domains many of which regularly reappear in the different proteins. These have often a complex secondary structure, maintained by multiple disulfide bonds. Whereas by denaturing and then carefully renaturing, an approximation to the native structure may be obtained using prokaryotic expression systems, and the best that may be expected is that a small percentage of the protein folds into such a conformation. Second, most ECM proteins are at least to some extent glycosylated and often heavily so, and the use of the mammalian system offers the best approximation to the sugar structures present in the native form of the molecule.
authors.affiliation: University of Cologne, Cologne, Germany.
authors.forename: Neil
authors.initials: N
authors.lastname: Smyth
authors.forename: Uwe
authors.initials: U
authors.lastname: Odenthal
authors.forename: Barbara
authors.initials: B
authors.lastname: Merkl
authors.forename: Mats
authors.initials: M
authors.lastname: Paulsson
mesh_terms: D002460:Cell Line; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D009842:Oligopeptides; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: C099517:Ala-Trp-Arg-His-Pro-Gln-Phe-Gly-Gly; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D009842:Oligopeptides; D011994:Recombinant Proteins