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Protein NMR pp 1–42Cite as

Assignment of Protein NMR Spectra Using Heteronuclear NMR—A Tutorial

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Part of the book series: Biological Magnetic Resonance ((BIMR,volume 32))

Abstract

The assignment of resonances in the complex nuclear magnetic resonance (NMR) spectrum of a protein is the first step in any NMR study of protein structure, function or dynamics. This chapter aims to provide a tutorial on protein NMR resonance assignment. Two approaches to the assignment are commonly used: the triple resonance methodology, which uses a suite of three-dimensional (3D) 13C/15N/1H NMR experiments, relies on through-bond 1J and 2J intra- and interresidue spin–spin couplings that are observed in 13C–15N double-labelled proteins; and the sequential assignment methodology, which can be applied to unlabelled or 15N single-labelled proteins, relies on through-bond total correlation spectroscopy (TOCSY) data to identify spin systems and through-space nuclear Overhauser effect (NOE) data to establish connections between neighbouring amino acid residues. This chapter describes both the methodologies for protein NMR resonance assignment. Examples of how sequence specific resonance assignments can be obtained using a suite of 2D and 3D NMR experiments are presented and suggestions on how overlap problems can be overcome are included.

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  1. Department of Biochemistry, University of Oxford, OX1 3QU, Oxford, UK

    Christina Redfield

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  1. Christina Redfield
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Correspondence to Christina Redfield .

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  1. Department of Chemistry and Biochemistry, University of Denver, Denver, Colorado, USA

    Lawrence Berliner

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Redfield, C. (2015). Assignment of Protein NMR Spectra Using Heteronuclear NMR—A Tutorial. In: Berliner, L. (eds) Protein NMR. Biological Magnetic Resonance, vol 32. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-7621-5_1

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