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    Rapid Distinction of Intracellular and Extracellular Proteins Using NMR Diffusion Measurements

    Waudby, Christopher A. and Mantle, M.D. and Cabrita, L.D. and Gladden, L.F. and Dobson, C.M. and Christodoulou, John (2012) Rapid Distinction of Intracellular and Extracellular Proteins Using NMR Diffusion Measurements. Journal of the American Chemical Society 134 (28), pp. 11312-11315. ISSN 0002-7863.

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    Abstract

    In-cell NMR spectroscopy offers a unique opportunity to begin to investigate the structures, dynamics, and interactions of molecules within their functional environments. An essential aspect of this technique is to define whether observed signals are attributable to intracellular species rather than to components of the extracellular medium. We report here the results of NMR measurements of the diffusion behavior of proteins expressed within bacterial cells, and find that these experiments provide a rapid and nondestructive probe of localization within cells and can be used to determine the size of the confining compartment. We show that diffusion can also be exploited as an editing method to eliminate extracellular species from high-resolution multidimensional spectra, and should be applicable to a wide range of problems. This approach is demonstrated here for a number of protein systems, using both 15N and 13C (methyl-TROSY) based acquisition.

    Metadata

    Item Type: Article
    School: Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences
    Research Centres and Institutes: Structural Molecular Biology, Institute of (ISMB)
    Depositing User: Administrator
    Date Deposited: 16 May 2013 09:48
    Last Modified: 02 Aug 2023 17:04
    URI: https://eprints.bbk.ac.uk/id/eprint/6789

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