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Perdeuteration/Site-Specific Protonation Approaches for High-Molecular-Weight Proteins

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Protein NMR Techniques

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 278))

Abstract

Among the factors that limit the application of nuclear magnetic resonance (NMR) to biological macromolecules are increasing resonance overlap and fast transverse relaxation. Multidimensional NMR combined with 13C and 15N labeling has alleviated these problems temporarily; however, they resurface at molecular weight (mol wt) in excess of 30 kDa. Combined perdeuteration/site-specific protonation together with segmental labeling (see Chapter 4), transverse relaxation-optimized spectroscopy (TROSY) (see Chapter 5), and residual dipolar couplings (see Chapter 7) have all helped to dramatically extend the mol wt limit. This article describes some of the practical aspects of the combined perdeuteration/site-specific protonation approach, which has proved so useful in the global fold determination of large proteins.

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Authors and Affiliations

  1. Department of Biological Sciences, Imperial College of Science, Technology and Medicine, London, UK

    Stephen Matthews

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  1. Stephen Matthews
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Editors and Affiliations

  1. Department of Biochemistry, University of Oxford, Oxford, UK

    A. Kristina Downing

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© 2004 Humana Press Inc., Totowa, NJ

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Matthews, S. (2004). Perdeuteration/Site-Specific Protonation Approaches for High-Molecular-Weight Proteins. In: Downing, A.K. (eds) Protein NMR Techniques. Methods in Molecular Biology™, vol 278. Humana Press. https://doi.org/10.1385/1-59259-809-9:035

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  • DOI: https://doi.org/10.1385/1-59259-809-9:035

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-246-9

  • Online ISBN: 978-1-59259-809-0

  • eBook Packages: Springer Protocols

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