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

Chu Kong Liew, Roland Gamsjaeger, Robyn E. Mansfield, Joel P. Mackay

    Research output: Contribution to journalArticlepeer-review

    9 Citations (Scopus)

    Abstract

    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 screening protein constructs for suitability as targets for structural studies.
    Original languageEnglish
    Pages (from-to)1630-1635
    Number of pages6
    JournalProtein Science
    Volume17
    Issue number9
    DOIs
    Publication statusPublished - 2008

    Fingerprint

    Dive into the research topics of 'NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins'. Together they form a unique fingerprint.

    Cite this