A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins.
Journal Article (Journal Article)
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.
Full Text
Duke Authors
Cited Authors
- Zhou, P; Lugovskoy, AA; Wagner, G
Published Date
- May 2001
Published In
Volume / Issue
- 20 / 1
Start / End Page
- 11 - 14
PubMed ID
- 11430750
International Standard Serial Number (ISSN)
- 0925-2738
Digital Object Identifier (DOI)
- 10.1023/a:1011258906244
Language
- eng
Conference Location
- Netherlands