Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

α-Synuclein occurs physiologically as a helically folded tetramer that resists aggregation

Abstract

Parkinson’s disease is the second most common neurodegenerative disorder1,2. Growing evidence indicates a causative role of misfolded forms of the protein α-synuclein in the pathogenesis of Parkinson’s disease3,4. Intraneuronal aggregates of α-synuclein occur in Lewy bodies and Lewy neurites5, the cytopathological hallmarks of Parkinson’s disease and related disorders called synucleinopathies4. α-Synuclein has long been defined as a ‘natively unfolded’ monomer of about 14 kDa (ref. 6) that is believed to acquire α-helical secondary structure only upon binding to lipid vesicles7. This concept derives from the widespread use of recombinant bacterial expression protocols for in vitro studies, and of overexpression, sample heating and/or denaturing gels for cell culture and tissue studies. In contrast, we report that endogenous α-synuclein isolated and analysed under non-denaturing conditions from neuronal and non-neuronal cell lines, brain tissue and living human cells occurs in large part as a folded tetramer of about 58 kDa. Several methods, including analytical ultracentrifugation, scanning transmission electron microscopy and in vitro cell crosslinking confirmed the occurrence of the tetramer. Native, cell-derived α-synuclein showed α-helical structure without lipid addition and had much greater lipid-binding capacity than the recombinant α-synuclein studied heretofore. Whereas recombinantly expressed monomers readily aggregated into amyloid-like fibrils in vitro, native human tetramers underwent little or no amyloid-like aggregation. On the basis of these findings, we propose that destabilization of the helically folded tetramer precedes α-synuclein misfolding and aggregation in Parkinson’s disease and other human synucleinopathies, and that small molecules that stabilize the physiological tetramer could reduce α-synuclein pathogenicity.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Western blot analysis of lysates of M17D, HeLa, HEK293 and COS-7 cells, mouse cortex and human RBCs probed for endogenous α-synuclein.
Figure 2: Sizing analyses of α-synuclein from human RBCs.
Figure 3: Comparative analyses of native (cell-derived) and bacterial α-synuclein.

References

  1. Obeso, J. A. et al. Missing pieces in the Parkinson’s disease puzzle. Nature Med. 16, 653–661 (2010)

    CAS  Article  Google Scholar 

  2. Gupta, A., Dawson, V. L. & Dawson, T. M. What causes cell death in Parkinson’s disease? Ann. Neurol. 64, S3–S15 (2008)

    CAS  Article  Google Scholar 

  3. Winklhofer, K. F., Tatzelt, J. & Haass, C. The two faces of protein misfolding: gain- and loss-of-function in neurodegenerative diseases. EMBO J. 27, 336–349 (2008)

    CAS  Article  Google Scholar 

  4. Tong, J. et al. Brain α-synuclein accumulation in multiple system atrophy, Parkinson’s disease and progressive supranuclear palsy: a comparative investigation. Brain 133, 172–188 (2010)

    Article  Google Scholar 

  5. Spillantini, M. G. et al. α-Synuclein in Lewy bodies. Nature 388, 839–840 (1997)

    ADS  CAS  Article  Google Scholar 

  6. Weinreb, P. H., Zhen, W., Poon, A. W., Conway, K. A. & Lansbury, P. T. J. NACP, a protein implicated in Alzheimer’s disease and learning, is natively unfolded. Biochemistry 35, 13709–13715 (1996)

    CAS  Article  Google Scholar 

  7. Davidson, W. S., Jonas, A., Clayton, D. F. & George, J. M. Stabilization of α-synuclein secondary structure upon binding to synthetic membranes. J. Biol. Chem. 273, 9443–9449 (1998)

    CAS  Article  Google Scholar 

  8. DeTure, M. et al. Missense tau mutations identified in FTDP-17 have a small effect on tau-microtubule interactions. Brain Res. 853, 5–14 (2000)

    CAS  Article  Google Scholar 

  9. Scherzer, C. R. et al. GATA transcription factors directly regulate the Parkinson’s disease-linked gene α-synuclein. Proc. Natl Acad. Sci. USA 105, 10907–10912 (2008)

    ADS  CAS  Article  Google Scholar 

  10. Wittig, I. & Schagger, H. Advantages and limitations of clear-native PAGE. Proteomics 5, 4338–4346 (2005)

    CAS  Article  Google Scholar 

  11. Osenkowski, P. et al. Cryoelectron microscopy structure of purified •-secretase at 12 Å resolution. J. Mol. Biol. 385, 642–652 (2009)

    CAS  Article  Google Scholar 

  12. Wall, J. S., Simon, M. N., Lin, B. Y. & Vinogradov, S. N. Mass mapping of large globin complexes by scanning transmission electron microscopy. Methods Enzymol. 436, 487–501 (2008)

    CAS  Article  Google Scholar 

  13. Beyer, K. Mechanistic aspects of Parkinson’s disease: α-synuclein and the biomembrane. Cell Biochem. Biophys. 47, 285–299 (2007)

    CAS  Article  Google Scholar 

  14. Chen, Y., Yang, J. T. & Martinez, H. M. Determination of the secondary structures of proteins by circular dichroism and optical rotatory dispersion. Biochemistry 11, 4120–4131 (1972)

    CAS  Article  Google Scholar 

  15. Sharon, R. et al. α-Synuclein occurs in lipid-rich high molecular weight complexes, binds fatty acids, and shows homology to the fatty acid-binding proteins. Proc. Natl Acad. Sci. USA 98, 9110–9115 (2001)

    ADS  CAS  Article  Google Scholar 

  16. Chen, P. S., Toribara, T. Y. & Warner, H. Microdetermination of phosphorus. Anal. Chem. 28, 1756–1758 (1956)

    CAS  Article  Google Scholar 

  17. Ko, L. W., Ko, H. H., Lin, W. L., Kulathingal, J. G. & Yen, S. H. Aggregates assembled from overexpression of wild-type α-synuclein are not toxic to human neuronal cells. J. Neuropathol. Exp. Neurol. 67, 1084–1096 (2008)

    CAS  Article  Google Scholar 

  18. McLean, P. J., Kawamata, H., Ribich, S. & Hyman, B. T. Membrane association and protein conformation of α-synuclein in intact neurons. Effect of Parkinson’s disease-linked mutations. J. Biol. Chem. 275, 8812–8816 (2000)

    CAS  Article  Google Scholar 

  19. Smith, D. P. et al. Formation of a high affinity lipid-binding intermediate during the early aggregation phase of α-synuclein. Biochemistry 47, 1425–1434 (2008)

    CAS  Article  Google Scholar 

  20. Tsika, E. et al. Distinct region-specific α-synuclein oligomers in A53T transgenic mice: implications for neurodegeneration. J. Neurosci. 30, 3409–3418 (2010)

    CAS  Article  Google Scholar 

  21. Klucken, J., Outeiro, T. F., Nguyen, P., McLean, P. J. & Hyman, B. T. Detection of novel intracellular α-synuclein oligomeric species by fluorescence lifetime imaging. FASEB J. 20, 2050–2057 (2006)

    CAS  Article  Google Scholar 

  22. Quintas, A., Saraiva, M. J. M. & Brito, R. M. M. The tetrameric protein transthyretin dissociates to a non-native monomer in solution. J. Biol. Chem. 274, 32943–32949 (1999)

    CAS  Article  Google Scholar 

  23. Connelly, S., Choi, S., Johnson, S. M., Kelly, J. W. & Wilson, I. A. Structure-based design of kinetic stabilizers that ameliorate the transthyretin amyloidoses. Curr. Opin. Struct. Biol. 20, 54–62 (2010)

    CAS  Article  Google Scholar 

  24. Lansbury, P. T. & Lashuel, H. A. A century-old debate on protein aggregation and neurodegeneration enters the clinic. Nature 443, 774–779 (2006)

    ADS  CAS  Article  Google Scholar 

Download references

Acknowledgements

Mass measurements were carried out at the Brookhaven National Laboratory STEM facility, a user facility supported by the US Department of Energy. We are grateful to D. Walker and J. Anderson (Elan Pharmaceuticals) for conducting mass spectrometry of our purified α-synuclein samples and for comments. We thank X. Simon and I. Perovic (Brandeis University) for their assistance with the AUC and phosphate analyses. Supported by NIH grants NS051318 and NS038375 (D.J.S.). We thank our colleagues at the Center for Neurologic Diseases for many discussions.

Author information

Authors and Affiliations

Authors

Contributions

All experiments were planned by T.B. and D.J.S. and conducted by T.B. and J.G.C. The manuscript was prepared by T.B. and D.J.S.

Corresponding author

Correspondence to Dennis J. Selkoe.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

The file contains Supplementary Figures 1-11 with legends. (PDF 848 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bartels, T., Choi, J. & Selkoe, D. α-Synuclein occurs physiologically as a helically folded tetramer that resists aggregation. Nature 477, 107–110 (2011). https://doi.org/10.1038/nature10324

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10324

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing