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Calcium-induced noncovalently linked tetramers of MRP8 and MRP14 are confirmed by electrospray ionization-mass analysis

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Journal of the American Society for Mass Spectrometry

Abstract

Proteins of the S100- family such as MRP8 (S100A8) and MRP14 (S100A9)—and its isoform MRP14*—show two calcium-binding sites (EF hands) per protein chain. MRP8, MRP14*, and MRP14, isolated from human granulocytes or monocytes, are known to form noncovalently associated complexes; the exact stoichiometries of these complexes in the presence of calcium are still controversially discussed in the literature. The present electrospray ionization-mass spectrometry (ESI-MS) study shows that MRP8, MRP14*, and MRP14 exist as heterodimers MRP8/14* and MRP8/14, respectively, in the absence of calcium confirming both a recent nuclear magnetic resonance study and a biochemical study on this topic. Furthermore, this ESI-MS study confirms the previously published matrix-assisted laser desorption ionization (MALDI)-MS study, which states that the MRP8/14* and MRP8/14 heterodimeric complexes tetramerize to heterotetramers (MRP8/14*)2, (MRP8/14*)(MRP8/14), and (MRP8/14)2, respectively, in the presence of calcium. The number of Ca2+ ions bound to the individual tetramer is determined to be eight for nonphosphorylated fractions; this is in agreement with the previously reported MALDI study on these fractions. About 1.2 Ca2+ ions more are bound to the phosphorylated form; it is speculated that the additional Ca2+ ions are bound to the phosphate groups in the tetramers. This study is, therefore, convincing proof of the reliability of MALDI-MS in studying noncovalent protein-protein interactions.

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Authors and Affiliations

  1. Laboratoire de Spectrométrie de Masse Bio-Organique (LSMBO), Université Louis Pasteur, Strasbourg, France

    Kerstin Strupat, Hélène Rogniaux & Alain Van Dorsselaer

  2. Institut für Experimentelle Dermatologie, Westfälische Wilhelms-Universität Münster, Münster, Germany

    Johannes Roth & Thomas Vogl

Authors
  1. Kerstin Strupat
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  2. Hélène Rogniaux
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  3. Alain Van Dorsselaer
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  4. Johannes Roth
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  5. Thomas Vogl
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Correspondence to Kerstin Strupat.

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Strupat, K., Rogniaux, H., Van Dorsselaer, A. et al. Calcium-induced noncovalently linked tetramers of MRP8 and MRP14 are confirmed by electrospray ionization-mass analysis. J. Am. Soc. Spectrom. 11, 780–788 (2000). https://doi.org/10.1016/S1044-0305(00)00150-1

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  • DOI: https://doi.org/10.1016/S1044-0305(00)00150-1

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