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Local cooperativity in the unfolding of an amyloidogenic variant of human lysozyme

Nature Structural Biology volume 9, pages 308–315 (2002)Cite this article

Abstract

Hydrogen exchange experiments monitored by NMR and mass spectrometry reveal that the amyloidogenic D67H mutation in human lysozyme significantly reduces the stability of the β-domain and the adjacent C-helix in the native structure. In addition, mass spectrometric data reveal that transient unfolding of these regions occurs with a high degree of cooperativity. This behavior results in the occasional population of a partially structured intermediate in which the three α-helices that form the core of the α-domain still have native-like structure, whereas the β-domain and C-helix are simultaneously substantially unfolded. This finding suggests that the extensive intermolecular interactions that will be possible in such a species are likely to initiate the aggregation events that ultimately lead to the formation of the well-defined fibrillar structures observed in the tissues of patients carrying this mutation in the lysozyme gene.

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Figure 1: 15N-1H HSQC NMR spectra of the wild type and D67H variant lysozymes.
Figure 2: Comparison of the protection factors for the D67H variant and wild type lysozyme.
Figure 3: Acceleration of hydrogen exchange rates in a substructure of the lysozyme variant.
Figure 4: Electrospray mass spectra of wild type and variant lysozymes demonstrating the unfolding dynamics.
Figure 5: Hydrogen exchange protection as a function of time.
Figure 6: Electrospray mass spectra of the partially exchanged peptide fragments of the D67H variant.

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Acknowledgements

D.C. was supported by a fellowship from the European Community, C.R. is supported by a BBSRC Advanced Research Fellowship, and C.V.R. and M.S. hold Royal Society University Research Fellowships. This work is in part a contribution from the Oxford Centre for Molecular Sciences, which is supported by EPSRC, BBSRC and MRC. The research of C.M.D. is also supported in part by a programme grant from the Wellcome Trust.

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Author notes

  1. Carol V. Robinson & Christopher M. Dobson

    Present address: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK

  2. Denis Canet

    Present address: Oxford Glycosciences UK Ltd., The Forum, 86 Milton Park, Abingdon, OX14 RY, UK

  3. David B. Archer

    Present address: School of Life and Environmental Sciences, University of Nottingham, University Park, Nottingham, NG7 2RD, UK

Authors and Affiliations

  1. Oxford Centre for Molecular Sciences, New Chemistry Laboratory, University of Oxford, South Parks Road, Oxford, OX1 3QH, UK

    Denis Canet, Alexander M. Last, Paula Tito, Christina Redfield, Carol V. Robinson & Christopher M. Dobson

  2. Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1EW, UK

    Margaret Sunde

  3. Institute of Food Research, Norwich Research Park, Colney, NR4 7UA, Norwich, UK

    Andrew Spencer

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Canet, D., Last, A., Tito, P. et al. Local cooperativity in the unfolding of an amyloidogenic variant of human lysozyme. Nat Struct Mol Biol 9, 308–315 (2002). https://doi.org/10.1038/nsb768

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