Eukaryotic Expression and Purification of Recombinant Extracellular Matrix Proteins Carrying the Strep II Tag

  • Neil Smyth
  • Uwe Odenthal
  • Barbara Merkl
  • Mats Paulsson
Part of the Methods in Molecular Biology™ book series (MIMB, volume 139)

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 (1) 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, 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.

Keywords

Recombinant Protein Column Volume Prokaryotic Expression System Episomal Plasmid Domain Border 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 2000

Authors and Affiliations

  • Neil Smyth
    • 1
  • Uwe Odenthal
    • 2
  • Barbara Merkl
    • 3
  • Mats Paulsson
    • 3
  1. 1.Institute for BiochemistryUniversity of CologneCologne
  2. 2.Institute for BiochemistryUniversity of OxfordOxfordUK
  3. 3.Institute for BiochemistryUniversity of CologneCologneGermany

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