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The extracellular chaperone clusterin sequesters oligomeric forms of the amyloid-β1−40 peptide

Abstract

In recent genome-wide association studies, the extracellular chaperone protein, clusterin, has been identified as a newly-discovered risk factor in Alzheimer's disease. We have examined the interactions between human clusterin and the Alzheimer's disease–associated amyloid-β1−40 peptide (Aβ1−40), which is prone to aggregate into an ensemble of oligomeric intermediates implicated in both the proliferation of amyloid fibrils and in neuronal toxicity. Using highly sensitive single-molecule fluorescence methods, we have found that Aβ1−40 forms a heterogeneous distribution of small oligomers (from dimers to 50-mers), all of which interact with clusterin to form long-lived, stable complexes. Consequently, clusterin is able to influence both the aggregation and disaggregation of Aβ1−40 by sequestration of the Aβ oligomers. These results not only elucidate the protective role of clusterin but also provide a molecular basis for the genetic link between clusterin and Alzheimer's disease.

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Figure 1: Bulk and single-molecule studies of Aβ1−40.
Figure 2: The effects of clusterin on the aggregation of Aβ1−40.
Figure 3: The effects of clusterin on the disaggregation of Aβ1−40 fibrils.

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Acknowledgements

P.N. is supported by the Marshall Aid Commemoration Commission (Marshall Scholarship) and the National Science Foundation (Graduate Research Fellowship). A.O. is supported by an European Reintegration Grant from the European Union FP7. R.W.C. acknowledges a Research Fellowship from Christ's College. B.B. is supported by an Alzheimer's Research Trust Fellowship. S.H. is supported by a Biotechnology and Biological Sciences Research Council Fellowship. K.A.G. is supported by fellowships from the Engineering and Physical Sciences Research Council and Studienstiftung des deutschen Volkes. S.M. is supported by a Royal Society Dorothy Hodgkin Fellowship. M.R.W. acknowledges the support of the Australian Research Council (DP0773555 and DP0984341). D.K. and C.M.D. are supported by the Wellcome Trust and the Medical Research Council. D.K. is supported by the Augustus Newman Foundation. Additionally, we would like to thank J. McColl and P. Dunne for advice on using the TIRFM instrument, S. Shammas and F. Newby for help with peptide and protein preparation, and T. Jahn, N. Cremades, L. Luheshi, T. Knowles, S. Cohen, A. Chen and J. Kumita for helpful discussions.

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Contributions

P.N., A.O., S.M., M.R.W., C.M.D. and D.K. designed the experiments. P.N. conducted the cTCCD experiments. P.N., A.O. and R.W.C. refined analysis methods. A.O. and R.W.C. developed instrumentation. R.W.C. wrote the analysis software, and designed, built and calibrated the scanning stage used for cTCCD experiments. P.N. and B.B. conducted the bulk scale experiments. P.N. and K.A.G. conducted the TIRFM experiments. P.N., B.B., K.A.G. and A.O. analyzed the data. S.H. labeled the clusterin that was purified and provided by M.R.W. All authors discussed and interpreted results and contributed to the writing of the manuscript.

Corresponding authors

Correspondence to Mark R Wilson or Christopher M Dobson or David Klenerman.

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Narayan, P., Orte, A., Clarke, R. et al. The extracellular chaperone clusterin sequesters oligomeric forms of the amyloid-β1−40 peptide. Nat Struct Mol Biol 19, 79–83 (2012). https://doi.org/10.1038/nsmb.2191

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