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Ion Mobility-Mass Spectrometry to Evaluate the Effects of Protein Modification or Small Molecule Binding on Protein Dynamics

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Ion Mobility-Mass Spectrometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2084))

Abstract

Ion mobility-mass spectrometry (IM-MS) of intact protein complexes under native conditions is a powerful tool for the analysis of protein complexes and protein–ligand interactions, permitting insight into ligand-induced changes in protein conformation. Here we describe a procedure for analyzing the effects of phosphorylation and/or inhibitor binding on protein kinase conformational flexibility using Protein Kinase A (PKA) as a model system. By calculating the protein collision cross section (CCS) before and after inhibitor binding, and additionally by performing collision-induced unfolding (CIU), we can establish the effects of protein modification or small molecule binding on protein dynamics.

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Author information

Authors and Affiliations

  1. Department of Biochemistry, Centre for Proteome Research, Institute of Integrative Biology, University of Liverpool, Liverpool, UK

    Lauren J. Tomlinson & Claire E. Eyers

Authors
  1. Lauren J. Tomlinson
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  2. Claire E. Eyers
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Corresponding author

Correspondence to Claire E. Eyers .

Editor information

Editors and Affiliations

  1. Department of Medicine and Surgery, University of Milano-Bicocca, Vedano al Lambro, Monza e Brianza, Italy

    Giuseppe Paglia

  2. Georgetown University, Washington, DC, USA

    Giuseppe Astarita

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Tomlinson, L.J., Eyers, C.E. (2020). Ion Mobility-Mass Spectrometry to Evaluate the Effects of Protein Modification or Small Molecule Binding on Protein Dynamics. In: Paglia, G., Astarita, G. (eds) Ion Mobility-Mass Spectrometry . Methods in Molecular Biology, vol 2084. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0030-6_11

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  • DOI: https://doi.org/10.1007/978-1-0716-0030-6_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0029-0

  • Online ISBN: 978-1-0716-0030-6

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