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Conformational changes in IgE contribute to its uniquely slow dissociation rate from receptor FcɛRI

Nature Structural & Molecular Biology volume 18, pages 571–576 (2011)Cite this article

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Abstract

Among antibody classes, IgE has a uniquely slow dissociation rate from, and high affinity for, its cell surface receptor FcɛRI. We show the structural basis for these key determinants of the ability of IgE to mediate allergic hypersensitivity through the 3.4-Å-resolution crystal structure of human IgE-Fc (consisting of the Cɛ2, Cɛ3 and Cɛ4 domains) bound to the extracellular domains of the FcɛRI α chain. Comparison with the structure of free IgE-Fc (reported here at a resolution of 1.9 Å) shows that the antibody, which has a compact, bent structure before receptor engagement, becomes even more acutely bent in the complex. Thermodynamic analysis indicates that the interaction is entropically driven, which explains how the noncontacting Cɛ2 domains, in place of the flexible hinge region of IgG antibodies, contribute together with the conformational changes to the unique binding properties of IgE.

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Figure 1: Overall structure of the IgE-Fc–sFcɛRIα complex, in two approximately orthogonal views.
Figure 2: Closure of an interdomain cleft in IgE-Fc upon receptor binding.
Figure 3: Interactions at the two subsites.
Figure 4: Thermodynamics of the IgE-FcɛRI interaction.

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Acknowledgements

We thank N. Rahman for assistance with receptor protein production (Oxford Protein Production Facility), T. Walter for technical advice on crystallization of the complex, and K. Harlos and C. Siebold (Division of Structural Biology, Wellcome Trust Centre for Human Genetics) for X-ray data collection of the complex, together with beamline support staff at European Synchrotron Radiation Facility. This work was also carried out with the support of the Diamond Light Source. The work was funded by Asthma UK and the UK Medical Research Council.

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Author notes

  1. Sarah C Bagby & James Hunt

    Present address: Present addresses: University of California, Department of Earth Science, Marine Science Institute, Santa Barbara, California, USA (S.C.B.); Novartis Horsham Research Centre, Horsham, UK (J.H.).,

  2. Mary D Holdom and Anna M Davies: These authors contributed equally to this work.

Authors and Affiliations

  1. Randall Division of Cell and Molecular Biophysics, King's College London, London, UK

    Mary D Holdom, Anna M Davies, Balvinder Dhaliwal, Enrico Girardi, James Hunt, Hannah J Gould, Andrew J Beavil, James M McDonnell & Brian J Sutton

  2. Medical Research Council & Asthma UK Centre in Allergic Mechanisms of Asthma, London, UK

    Mary D Holdom, Anna M Davies, Balvinder Dhaliwal, Enrico Girardi, Hannah J Gould, Andrew J Beavil, James M McDonnell & Brian J Sutton

  3. Division of Structural Biology, University of Oxford, The Oxford Protein Production Facility, Oxford, UK

    Joanne E Nettleship & Ray J Owens

  4. Oxford Protein Production Facility UK, The Research Complex at Harwell, Rutherford Appleton Laboratory, Harwell Science & Innovation Centre, Harwell, UK

    Joanne E Nettleship & Ray J Owens

  5. Department of Biochemistry, University of Oxford, Laboratory of Molecular Biophysics, Oxford, UK

    Sarah C Bagby

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Contributions

M.D.H. and A.M.D. carried out the crystallographic analysis of the complex, and B.D. the crystallographic analysis of IgE-Fc; M.D.H., J.E.N., J.H., A.J.B. and R.J.O. produced the proteins; S.C.B. and J.M.M. carried out the thermodynamic analysis; E.G. contributed to the analysis of the conformational changes; H.J.G., A.J.B. and B.J.S. planned and directed the project; M.D.H., A.M.D., J.M.M. and B.J.S. wrote the paper.

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Correspondence to Brian J Sutton.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 (PDF 706 kb)

Supplementary Video 1

Overall structure of the IgE-Fc/sFcɛRIα complex. This video shows the proposed orientation of the complex in relation to the cell membrane and highlights the acute bend in IgE-Fc, with the Cɛ2 domains packed against the Cɛ3 and Cɛ4 domains. The complex is rotated by 90° clockwise, 180° anti-clockwise, and 90° clockwise about an axis orthogonal to the cell membrane. IgE-Fc chains A and B are colored in green and purple respectively, and sFcɛRIα in yellow. (MOV 2938 kb)

Supplementary Video 2

Conformational change in IgE-Fc. This video demonstrates the conformational change that takes place in IgE-Fc upon sFcɛRIα binding. The video shows the same conformational change from four views, each 90° apart. The change is demonstrated by morphing the free structure of IgE-Fc into that of the receptor-bound form, and then back to the free form. IgE-Fc chains A and B are colored in green and purple, respectively. (MOV 11684 kb)

Supplementary Video 3

Conformational change in Cɛ2 and Cɛ3 domains on receptor binding. This video demonstrates the conformational change that takes place in IgE-Fc upon sFcɛRIα binding (with emphasis on the Cɛ2 domains), and shows how the Cɛ2A, Cɛ2B and Cɛ3A domains move together as a rigid unit. The video also shows the small conformational change within Cɛ3B. The free structure of IgE-Fc is first shown, and then morphed into the receptor-bound form. The receptor is briefly displayed, after which IgE-Fc is morphed back to the free form. IgE-Fc chains A and B are colored in green and purple respectively, and sFcɛRIα in yellow. (MOV 2149 kb)

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Holdom, M., Davies, A., Nettleship, J. et al. Conformational changes in IgE contribute to its uniquely slow dissociation rate from receptor FcɛRI. Nat Struct Mol Biol 18, 571–576 (2011). https://doi.org/10.1038/nsmb.2044

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