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For the Record

NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins

University of Sydney

(RECEIVED February 17, 2008; ACCEPTED April 23, 2008)

Glutathione-S-transferase (GST) fusion proteins are used extensively for structural, biochemical and functional analyses. Although the conformation of the target protein is of critical importance, confirmation of the folded state of the target is often not undertaken, or is cumbersome because of the requirement to first remove the GST tag. Here, we demonstrate that it is possible to record conventional ¹⁵N-HSQC NMR spectra of small GST-fusion proteins and that the observed signals arise almost exclusively from the target protein. This approach constitutes a rapid and straightforward means of assessing the conformation of a GST-fusion protein without having to cleave the GST, and should prove valuable both to biochemists seeking to check the conformation of their proteins prior to functional studies and to structural biologists who are screening protein constructs for their suitability as targets for structural studies.

Keywords: Protein Structure/Folding; Structure/function studies; NMR Spectroscopy; Heteronuclear NMR; Genomics - Structural

¹ E-mail: ckliew{at}mmb.usyd.edu.au

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