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A 31-residue peptide induces aggregation of tau's microtubule-binding region in cells

Nature Chemistry volume 9, pages 874–881 (2017)Cite this article

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Abstract

The self-propagation of misfolded conformations of tau underlies neurodegenerative diseases, including Alzheimer's. There is considerable interest in discovering the minimal sequence and active conformational nucleus that defines this self-propagating event. The microtubule-binding region, spanning residues 244–372, reproduces much of the aggregation behaviour of tau in cells and animal models. Further dissection of the amyloid-forming region to a hexapeptide from the third microtubule-binding repeat resulted in a peptide that rapidly forms fibrils in vitro. We show that this peptide lacks the ability to seed aggregation of tau244–372 in cells. However, as the hexapeptide is gradually extended to 31 residues, the peptides aggregate more slowly and gain potent activity to induce aggregation of tau244–372 in cells. X-ray fibre diffraction, hydrogen–deuterium exchange and solid-state NMR studies map the beta-forming region to a 25-residue sequence. Thus, the nucleus for self-propagating aggregation of tau244–372 in cells is packaged in a remarkably small peptide.

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Figure 1: Amyloid formation of R3-derived peptides and transmission to HEK(tau-RD(LM)-YFP) cells.
Figure 2: Amyloid formation and self-propagation of covalent variation of R3.
Figure 3: Fluorescence emission fingerprinting of R3 amyloids.
Figure 4: Structural investigations reveal a common parallel, stacked beta-sheet architecture in R3 amyloids.
Figure 5: Solid-state NMR spectra of R3SH fibrils show a homogenous, predominantly beta-strand structure.

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Acknowledgements

J.S., W.F.D., H.W., M.N. and S.B.P. are supported by grants from the National Institutes of Health (AG002132, AG031220, GM054616 and HL007731). J.S. and M.N. were supported by an award from the Glenn Foundation. J.S. is a recipient of the New Investigator Research Grant (2015-NIRG-339935) from the Alzheimer's Association. F.G. and J.R. are supported by a grant from the National Institutes of Health (GM104605). This work was also supported in part by the UCSF Research Resource Program Shared Equipment Award funded by the UCSF Office of Research, Daiichi Sankyo, the Henry M. Jackson Foundation, the Sherman Fairchild Foundation, and by a gift from the Rainwater Charitable Foundation.

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Author notes

  1. Jan Stöhr, Haifan Wu and Mimi Nick: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute for Neurodegenerative Diseases, University of California, San Francisco, 94143, California, USA

    Jan Stöhr, Carlo Condello, Noah Johnson, Srabasti Acharya, Julia Becker, Kathleen Robinson, Stanley B. Prusiner & William F. DeGrado

  2. Department of Neurology, University of California, San Francisco, 94143, California, USA

    Jan Stöhr, Carlo Condello & Stanley B. Prusiner

  3. Department of Pharmaceutical Chemistry, University of California, San Francisco, 94143, California, USA

    Haifan Wu, Mimi Nick, Yibing Wu, Manasi Bhate, Thomas Lemmin, Mark J. S. Kelly & William F. DeGrado

  4. Department of Chemistry, University of Pennsylvania, Philadelphia, 19104, Pennsylvania, USA

    Jeffrey Rodgers & Feng Gai

  5. Department of Biological Sciences and Center for Structural Biology, Vanderbilt University, Nashville, 37235, Tennessee, USA

    Gerald Stubbs

  6. Department of Biochemistry and Biophysics, University of California, San Francisco, 94143, California, USA

    Stanley B. Prusiner

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  1. Jan Stöhr
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Contributions

J.S., W.F.D. and S.B.P. conceived and designed the experiments; J.S., M.N., H.W., M.B., C.C., N.J., J.R., T.L., S.A., J.B., M.J.S.K. and K.R. performed the experiments; J.S., M.N., H.W.Y.W., M.B. and C.C analysed data; J.S., W.F.D. and S.B.P. co-wrote the paper; J.S., M.N. and H.W. contributed equally to this work.

Corresponding authors

Correspondence to Jan Stöhr or William F. DeGrado.

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Competing interests

UCSF is conducting research sponsored by Daiichi Sankyo (Tokyo, Japan) to develop diagnostics and therapeutics for tau. Currently, there is no direct relationship between these agreements and this Article.

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Stöhr, J., Wu, H., Nick, M. et al. A 31-residue peptide induces aggregation of tau's microtubule-binding region in cells. Nature Chem 9, 874–881 (2017). https://doi.org/10.1038/nchem.2754

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