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    An integrative approach combining ion mobility mass spectrometry, X-ray crystallography and NMR spectroscopy to study the conformational dynamics of α1-antitrypsin upon ligand binding

    Nyon, M.P. and Prentice, T. and Day, J. and Kirkpatrick, J. and Sivalingam, G.N. and Lévy, Géraldine R. and Haq, I. and Irving, J. and Lomas, D.A. and Christodoulou, John and Gooptu, Bibek and Thalassinos, Konstantinos (2015) An integrative approach combining ion mobility mass spectrometry, X-ray crystallography and NMR spectroscopy to study the conformational dynamics of α1-antitrypsin upon ligand binding. Protein Science 24 (8), pp. 1301-1312. ISSN 0961-8368.

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    Abstract

    Native mass spectrometry (MS) methods permit the study of multiple protein species within solution equilibria, whilst ion mobility (IM)-MS can report on conformational behaviour of specific states. We used IM-MS to study a conformationally labile protein (α1-antitrypsin) that undergoes pathological polymerisation in the context of point mutations. The folded, native state of the Z variant remains highly polymerogenic in physiological conditions, despite only minor thermodynamic destabilisation relative to the wild-type variant. Various data implicate kinetic instability (conformational lability within a native state ensemble) as the basis of Z α1-antitrypsin polymerogenicity. We show the ability of IM-MS to track such disease-relevant conformational behaviour in detail by studying the effects of peptide binding on α1-antitrypsin conformation and dynamics. IM-MS is therefore an ideal platform for the screening of compounds that result in therapeutically-beneficial kinetic stabilisation of native α1-antitrypsin. Our findings are confirmed with high resolution X-ray crystallographic and NMR spectroscopic studies of the same event, which together dissect structural changes from dynamic effects caused by peptide binding at a residue specific level. IM-MS methods therefore have great potential for further study of biologically-relevant thermodynamic and kinetic instability of proteins and provide rapid and multidimensional characterisation of ligand interactions of therapeutic interest. This article is protected by copyright. All rights reserved.

    Metadata

    Item Type: Article
    Additional Information: *Bibek Gooptu credited as joint senior/corresponding author* This is the peer reviewed version of the article, which has been published in final form at the link cited. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    Research Centre: Structural Molecular Biology, Institute of (ISMB)
    Depositing User: Administrator
    Date Deposited: 23 Jun 2015 12:19
    Last Modified: 09 Dec 2019 02:09
    URI: http://eprints.bbk.ac.uk/id/eprint/12412

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