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Electrospray ionization mass spectrometry and hydrogen/deuterium exchange for probing the interaction of calmodulin with calcium

  • Focus: H/D Exchange Of Proteins In Solution
  • Published:
Journal of the American Society for Mass Spectrometry

Abstract

The extent of H/D exchange of the protein calmodulin in solution was monitored by mass spectrometry following electrospray ionization (ESI) of the protein. In the absence of Ca2+, approximately 115 protons are exchanged for deuteriums after 60 min. As the calmodulin is titrated with Ca2+, the extent of exchange decreases significantly (i.e., by 24 protons), indicating Ca2+-induced folding of the protein to a tighter, less solvent-accessible form. The extent of H/D exchange ceases to decrease when the amount of added Ca2+ is sufficient to convert greater than 80% of the calmodulin to a form bound by four calcium ions. Lysozyme, a protein of similar molecular weight, does not show a significant decrease in the extent of H/D exchange as it binds to Ca2+, indicating that the changes in H/D exchange for calmodulin reflect tertiary structural change that occur upon binding with Ca2+.

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Authors and Affiliations

  1. Department of Chemistry, Washington University, St. Louis, Missouri, USA

    Olga Nemirovskiy, Daryl E. Giblin & Michael L. Gross

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  1. Olga Nemirovskiy
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  2. Daryl E. Giblin
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  3. Michael L. Gross
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Correspondence to Michael L. Gross.

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Nemirovskiy, O., Giblin, D.E. & Gross, M.L. Electrospray ionization mass spectrometry and hydrogen/deuterium exchange for probing the interaction of calmodulin with calcium. J Am Soc Mass Spectrom 10, 711–718 (1999). https://doi.org/10.1016/S1044-0305(99)00036-7

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  • DOI: https://doi.org/10.1016/S1044-0305(99)00036-7

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