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    Cell-free expression and selective isotope labelling in protein NMR

    Staunton, D. and Schlinkert, R. and Zanetti, Giulia and Colebrook, S.A. and Campbell, I.D. (2006) Cell-free expression and selective isotope labelling in protein NMR. Magnetic resonance in chemistry 44 (S1), S2-S9. ISSN 1097-458X.

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    Abstract

    Isotope labelling is a very powerful tool in NMR studies of proteins and has been employed in various ways for over 40 years. 15N and 13C incorporation, using recombinant expression systems, is now commonplace because heteronuclear experiments assist with the fundamental problems of peak resolution and assignment. The use of selective labelling for peak assignment has been restricted by the scrambling of isotope label through metabolic pathways within the expression host organism. The availability of efficient cell‐free expression systems with low levels of metabolic conversion allow the increasing use of selective isotope labelling as a tool in protein NMR. We describe two examples, one where a selective labelling scheme can identify backbone amide peaks from unassigned 1H15N HSQC and HNCO spectra of a 84 residue protein, and another where a specific backbone amide in a 198 residue construct of the ninth and tenth Type III repeats from human fibronectin can be labelled and rapidly identified using a simple HSQC experiment. Copyright © 2006 John Wiley & Sons, Ltd.

    Metadata

    Item Type: Article
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    Depositing User: Sarah Hall
    Date Deposited: 20 May 2019 11:18
    Last Modified: 20 May 2019 11:18
    URI: http://eprints.bbk.ac.uk/id/eprint/27573

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