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Nondenaturing Mass Spectrometry to Study Noncovalent Protein/Protein and Protein/Ligand Complexes: Technical Aspects and Application to the Determination of Binding Stoichiometries

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Functional Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 484))

Abstract

In the present chapter we detail how mass spectrometry (MS) can be used to characterize noncovalent complexes, especially multimeric proteins and protein/ligand complexes. This original application of MS, also called “supramolecular MS” or “nondenaturing MS,” appeared in the early 1990s and has continuously evolved since then. Nondenaturing MS is now fully integrated in structural biology programs and in drug discovery platforms. Indeed, appropriate sample preparation and fine tuning of the instrument make it possible to transfer weak assemblies without disruption from solution into the gas phase of the mass spectrometer. In this chapter we detail experimental conditions (sample preparation, optimization of instrumental parameters, etc.) required for the detection of noncovalent complexes by MS. We then focus on the type of information and accuracy that we get after interpreting electrospray ionization mass spectra obtained under nondenaturing conditions, with emphasis on the determination of the stoichiometry of protein/protein and protein/ligand complexes.

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Authors and Affiliations

  1. Laboratoire de Spectrométrie de Masse Bio-Organique, Institut Pluridisciplinaire Hubert Curien, UMR 7178 CNRS/Université Louis Pasteur, Strasbourg, France

    Sarah Sanglier, Cédric Atmanene, Guillaume Chevreux & Alain Van Dorsselaer

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  1. Sarah Sanglier
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  2. Cédric Atmanene
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  3. Guillaume Chevreux
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  4. Alain Van Dorsselaer
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Editor information

Editors and Affiliations

  1. Laboratoire de Bioinformatique et Génomique Intégratives, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France

    Julie D. Thompson

  2. Department of Protein Science Helmholtz Zentrum München, German Research Center for Environmental Health, Munich-Neuherberg, Germany

    Marius Ueffing

  3. LSMBO, ECPM, Institut Pluridisciplinaire Hubert Curien, Strasbourg, France

    Christine Schaeffer-Reiss

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© 2008 Humana Press, Totowa, NJ

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Sanglier, S., Atmanene, C., Chevreux, G., Van Dorsselaer, A. (2008). Nondenaturing Mass Spectrometry to Study Noncovalent Protein/Protein and Protein/Ligand Complexes: Technical Aspects and Application to the Determination of Binding Stoichiometries. In: Thompson, J.D., Ueffing, M., Schaeffer-Reiss, C. (eds) Functional Proteomics. Methods in Molecular Biology, vol 484. Humana Press. https://doi.org/10.1007/978-1-59745-398-1_15

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  • DOI: https://doi.org/10.1007/978-1-59745-398-1_15

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-971-0

  • Online ISBN: 978-1-59745-398-1

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