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Probing Backbone Dynamics with Hydrogen/Deuterium Exchange Mass Spectrometry

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Protein Dynamics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1084))

Abstract

Protein dynamics can be probed by the solution technique amide hydrogen/deuterium exchange. The exchange rate of hydrogen for deuterium along a peptide backbone is dependent on the extent of hydrogen bonding from secondary structure, accessibility by D2O, and protein motions. Both global and local conformational changes that alter bonding or structure will lead to changes in the amount of deuterium incorporated. The deuterium can be localized via pepsin digestion of the protein and quantified by electrospray ionization mass spectrometry through the mass shifts of the resulting peptides. The technique is emerging as an essential tool to study protein structure in solution due to the exceptional capability of examining both dynamic and structural changes related to protein function.

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Acknowledgement

This work was supported by NSF grant #0845273 to L.S.B.

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Authors and Affiliations

  1. Department of Chemistry, The University of Alabama, Tuscaloosa, AL, USA

    Harsimran Singh & Laura S. Busenlehner

Authors
  1. Harsimran Singh
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  2. Laura S. Busenlehner
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Editor information

Editors and Affiliations

  1. Department of Bioinformatics and Genomic, University of North Carolina at Charlott, Charlotte, North Carolina, USA

    Dennis R. Livesay

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Singh, H., Busenlehner, L.S. (2014). Probing Backbone Dynamics with Hydrogen/Deuterium Exchange Mass Spectrometry. In: Livesay, D. (eds) Protein Dynamics. Methods in Molecular Biology, vol 1084. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-658-0_5

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  • DOI: https://doi.org/10.1007/978-1-62703-658-0_5

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-657-3

  • Online ISBN: 978-1-62703-658-0

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