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Journal of Biomolecular NMR

, Volume 20, Issue 1, pp 11–14 | Cite as

A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins

  • Pei Zhou
  • Alexey A. Lugovskoy
  • Gerhard Wagner
Article

Abstract

Protein-fusion constructs have been used with great success for enhancing expression of soluble recombinant protein and as tags for affinity purification. Unfortunately the most popular tags, such as GST and MBP, are large, which hinders direct NMR studies of the fusion proteins. Cleavage of the fusion proteins often re-introduces problems with solubility and stability. Here we describe the use of N-terminally fused protein G (B1 domain) as a non-cleavable solubility-enhancement tag (SET) for structure determination of a dimeric protein complex. The SET enhances the solubility and stability of the fusion product dramatically while not interacting directly with the protein of interest. This approach can be used for structural characterization of poorly behaving protein systems, and would be especially useful for structural genomics studies.

protein G protein stabilization protein tag 

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Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Pei Zhou
  • Alexey A. Lugovskoy
  • Gerhard Wagner

There are no affiliations available

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