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Protein Structure, Stability, and Interactions pp 285–310Cite as

NMR Analysis of Native-State Protein Conformational Flexibility by Hydrogen Exchange

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 490))

Abstract

The rate of hydrogen exchange for the most protected amides of a protein is widely used to provide an estimate of global conformational stability by analyzing the exchange kinetics in the unfolded state in terms of model peptide exchange rates. The exchange behavior of the other amides of the protein which do not exchange via a global unfolding mechanism can provide insight into the smaller-scale conformational transitions that facilitate access to solvent as required for the exchange reaction. However, since the residual tertiary structure in the exchange-competent conformation can modulate the chemistry of the exchange reaction, equilibrium values estimated from normalization with model peptide rates are open to question. To overcome this limitation, the most robust approaches utilize differential analyses as a function of experimental variables such as denaturant concentration, temperature, pH, and mutational variation. Practical aspects of these various differential analysis techniques are considered with illustrations drawn from the literature.

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  1. Wadsworth Center, New York State Department of Health, and Department of Biomedical Sciences, University at Albany – SUNY, Albany, NY, USA

    Griselda Hernández & David M. LeMaster

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    Hernández, G., LeMaster, D.M. (2009). NMR Analysis of Native-State Protein Conformational Flexibility by Hydrogen Exchange. In: Shriver, J. (eds) Protein Structure, Stability, and Interactions. Methods in Molecular Biology, vol 490. Humana Press. https://doi.org/10.1007/978-1-59745-367-7_12

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