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H/D Exchange Centroid Monitoring is Insufficient to Show Differences in the Behavior of Protein States

  • Application Note
  • Published:
Journal of The American Society for Mass Spectrometry

Abstract

Differential hydrogen/deuterium exchange (H/DX) coupled with mass spectrometry (H/DX-MS) offers a rapid and sensitive characterization of changes in proteins following perturbations induced by changes in folding, ligand binding, oligomerization, and modification. The characterization of H/DX rates by software tools and automated data processing often relies on the centroid mass calculation and, thereby, the deuterium distribution in the mass spectra is neglected. Here we present an example demonstrating the clear limitation of using only a centroid approach to characterize the H/DX rate, in which the change in protein is not reflected as the difference in deuterium uptake based on centroid calculation.

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Acknowledgment

The authors thank Bruce Pascal from Professor Patrick R. Griffin’s lab at Scripps Florida for help with HDX data analysis and helpful discussion. The work was supported by grants from the National Institute of General Medical Sciences (8 P41 GM103422-35) of the NIH.

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Authors and Affiliations

  1. Department of Chemistry, Washington University in St. Louis, St. Louis, MO, 63130, USA

    Jun Zhang & Michael L. Gross

  2. Division of Nephrology and Hypertension, Department of Medicine, Mayo Clinic, Rochester, MN, 55905, USA

    Pradeep Ramachandran & Rajiv Kumar

  3. Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, 55905, USA

    Pradeep Ramachandran & Rajiv Kumar

Authors
  1. Jun Zhang
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  2. Pradeep Ramachandran
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  3. Rajiv Kumar
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  4. Michael L. Gross
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Correspondence to Michael L. Gross.

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Zhang, J., Ramachandran, P., Kumar, R. et al. H/D Exchange Centroid Monitoring is Insufficient to Show Differences in the Behavior of Protein States. J. Am. Soc. Mass Spectrom. 24, 450–453 (2013). https://doi.org/10.1007/s13361-012-0555-z

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  • DOI: https://doi.org/10.1007/s13361-012-0555-z

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