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The toxicity of antiprion antibodies is mediated by the flexible tail of the prion protein

Nature volume 501, pages 102–106 (2013)Cite this article

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Abstract

Prion infections cause lethal neurodegeneration. This process requires the cellular prion protein (PrPC; ref. 1), which contains a globular domain hinged to a long amino-proximal flexible tail2. Here we describe rapid neurotoxicity in mice and cerebellar organotypic cultured slices exposed to ligands targeting the α1 and α3 helices of the PrPC globular domain. Ligands included seven distinct monoclonal antibodies3, monovalent Fab1 fragments and recombinant single-chain variable fragment miniantibodies. Similar to prion infections4,5,6, the toxicity of globular domain ligands required neuronal PrPC, was exacerbated by PrPC overexpression, was associated with calpain activation and was antagonized by calpain inhibitors. Neurodegeneration was accompanied by a burst of reactive oxygen species, and was suppressed by antioxidants. Furthermore, genetic ablation of the superoxide-producing enzyme NOX2 (also known as CYBB) protected mice from globular domain ligand toxicity. We also found that neurotoxicity was prevented by deletions of the octapeptide repeats within the flexible tail. These deletions did not appreciably compromise globular domain antibody binding, suggesting that the flexible tail is required to transmit toxic signals that originate from the globular domain and trigger oxidative stress and calpain activation. Supporting this view, various octapeptide ligands were not only innocuous to both cerebellar organotypic cultured slices and mice, but also prevented the toxicity of globular domain ligands while not interfering with their binding. We conclude that PrPC consists of two functionally distinct modules, with the globular domain and the flexible tail exerting regulatory and executive functions, respectively. Octapeptide ligands also prolonged the life of mice expressing the toxic PrPC mutant7, PrP(Δ94–134), indicating that the flexible tail mediates toxicity in two distinct PrPC-related conditions. Flexible tail-mediated toxicity may conceivably play a role in further prion pathologies, such as familial Creutzfeldt-Jakob disease in humans bearing supernumerary octapeptides.

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Figure 1: Epitope specificity of the neurotoxicity mediated by antiprion antibodies.
Figure 2: Antibody-mediated toxicity in cerebellar slices does not require crosslinking or antibody effector functions.
Figure 3: Neuroprotection against globular domain ligand toxicity.
Figure 4: In vivo toxicity of antibodies binding to PrP.

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Protein Data Bank

Data deposits

X-ray crystallographic coordinates and structure factor files have been deposited in the RSCB Protein Data Bank (PDB) database under the accession code number 4H88.

Change history

  • 04 September 2013

    Minor changes were made to the Acknowledgements.

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Acknowledgements

We thank S. Izui, H. Monyer, D. Burton and G. Mallucci for reagents and mice, S. Schauer and the Functional Genomics Center Zurich for advice and help with affinity determinations, A. Steingötter, U. Ungethüm, M. Polymenidou, A. Lau and A. Keller for input, R. Moos, B. Sikorska, C. Tiberi, P. Schwarz, A. Varol, K. Arroyo and M. Delic for technical help. A.A. is the recipient of an Advanced Grant of the European Research Council and is supported by grants from the European Union (PRIORITY, LUPAS and NEURINOX), the Swiss National Foundation, the Foundation Alliance BioSecure, the Clinical Research Priority Program (KFSP) of the University of Zurich, and the Novartis Research Foundation. J.F. is supported by a career development award of the University of Zurich. Research support from PrioNet Canada and Alberta Prion Research Institute (APRI) for the work conducted in the Canadian laboratories is gratefully acknowledged. P.P.L. and A.A. are supported by Polish Swiss Research grant nr PSPB-062/2010. This paper is dedicated to the memory of Dr Marek Fischer, who created the tga20 mouse line.

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Author notes

  1. Tiziana Sonati, Regina R. Reimann and Jeppe Falsig: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Neuropathology, University Hospital Zurich, Schmelzbergstrasse 12, CH-8091 Zurich, Switzerland,

    Tiziana Sonati, Regina R. Reimann, Jeppe Falsig, Tracy O’Connor, Simone Hornemann, Sine Yaganoglu, Bei Li, Uli S. Herrmann & Adriano Aguzzi

  2. Department of Biochemistry, Faculty of Medicine & Dentistry, University of Alberta 474 Medical Sciences Building, Edmonton, Alberta T6G 2H7, Canada,

    Pravas Kumar Baral, Barbara Wieland, Mridula Swayampakula & Michael N. G. James

  3. Department of Agricultural, Food and Nutritional Science, University of Alberta, 410 Agriculture/Forestry Centre, Edmonton, Alberta T6G 2P5, Canada,

    Muhammad Hafizur Rahman, Dipankar Das & Nat Kav

  4. ETH Zurich, Physical Chemistry, ETH Hönggerberg, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich, Switzerland,

    Roland Riek

  5. Department of Molecular Pathology and Neuropathology, Laboratory of Electron Microscopy and Neuropathology, Medical University of Lodz, Czechoslowacka 8/10, 92-216 Lodz, Poland,

    Pawel P. Liberski

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Contributions

T.S., J.F. and A.A. conceived the study. Planning and execution were performed by T.S. with significant contributions from R.R.R., J.F. and additional contribution from T.O’C., S.H., S.Y., B.L. and U.S.H. MEMRI was established and performed by R.R.R.; electron microscopy was performed by P.P.L.; scFv were cloned and produced by B.W., M.S., M.H.R., D.D. and N.K.; X-ray crystallography was performed by P.K.B. and M.N.G.J.; and NMR experiments were performed by S.H. with additional contributions from R.R. T.S., J.F. and A.A. wrote the manuscript.

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Correspondence to Adriano Aguzzi.

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Supplementary information

Supplementary Information

This file contains Supplementary Tables 1-4 and Supplementary Figures 1- 15. (PDF 4156 kb)

Three-dimensional representation of the complex between rmPrP120-231 and F(ab)1POM1

Video derived from the structure of its crystal. The rotating image visualizes a discontinuous epitope on PrP with interactions involving both the β1-α1 loop and helix α3. (WMV 5916 kb)

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Sonati, T., Reimann, R., Falsig, J. et al. The toxicity of antiprion antibodies is mediated by the flexible tail of the prion protein. Nature 501, 102–106 (2013). https://doi.org/10.1038/nature12402

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