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Solid-state NMR structure of a pathogenic fibril of full-length human α-synuclein

Nature Structural & Molecular Biology volume 23, pages 409415 (2016) | Download Citation


Misfolded α-synuclein amyloid fibrils are the principal components of Lewy bodies and neurites, hallmarks of Parkinson's disease (PD). We present a high-resolution structure of an α-synuclein fibril, in a form that induces robust pathology in primary neuronal culture, determined by solid-state NMR spectroscopy and validated by EM and X-ray fiber diffraction. Over 200 unique long-range distance restraints define a consensus structure with common amyloid features including parallel, in-register β-sheets and hydrophobic-core residues, and with substantial complexity arising from diverse structural features including an intermolecular salt bridge, a glutamine ladder, close backbone interactions involving small residues, and several steric zippers stabilizing a new orthogonal Greek-key topology. These characteristics contribute to the robust propagation of this fibril form, as supported by the structural similarity of early-onset-PD mutants. The structure provides a framework for understanding the interactions of α-synuclein with other proteins and small molecules, to aid in PD diagnosis and treatment.

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This study was supported by the US National Institutes of Health (NIH) (grants R01-GM073770 to C.M.R., P50-NS053488 to V.M.Y.L. and P01-AG002132 to G.S.) and used SSNMR instrumentation procured with the support of grant S10-RR025037 (to C.M.R.) from the NIH National Center for Research Resources (NCRR). M.D.T., A.J.N. and A.M.B. were supported as members of the NIH Molecular Biophysics Training Grant at the University of Illinois at Urbana-Champaign (T32-GM008276), and D.J.C. is supported by grant T32-AG000255. J.M.C. was supported by a US National Science Foundation Graduate Research Fellowship. C.D.S. is supported by the Intramural Research Program of the Center for Information Technology at NIH. The authors thank J. Wall and B. Lin (Brookhaven National Laboratory) for STEM MPL sample preparation and data collection. The Brookhaven National Laboratory STEM was an NIH-supported Resource Center (P41-EB2181), and additional support was provided by the US Department of Energy (DOE), Office of Biological and Environmental Research. TEM images were collected at the Frederick Seitz Materials Research Laboratory Central Facilities, University of Illinois, which are partially supported by the DOE under grants DE-FG02-07ER46453 and DE-FG02-07ER46471. The Voyager-DE STR MALDI TOF mass spectrometer was purchased in part with a grant from the NIH NCRR (S10-RR011966). The Stanford Synchrotron Radiation Lightsource (SSRL) is a national user facility operated by Stanford University on behalf of the DOE, Office of Basic Energy Sciences. The SSRL Structural Molecular Biology Program is supported by the DOE and the NIH. The authors thank M. Tang (College of Staten Island) for helpful discussions.

Author information

Author notes

    • Marcus D Tuttle
    • , Andrew J Nieuwkoop
    • , Kathryn D Kloepper
    •  & William Wan

    Present addresses: Department of Chemistry, Yale University, New Haven, Connecticut, USA (M.D.T.), Leibniz-Institut Für Molekulare Pharmakologie, Berlin, Germany (A.J.N.), Department of Chemistry, Mercer University, Macon, Georgia, USA (K.D.K.) and Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany (W.W.).


  1. Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

    • Marcus D Tuttle
    • , Andrew J Nieuwkoop
    • , Deborah A Berthold
    • , Kathryn D Kloepper
    • , Joseph M Courtney
    • , Jae K Kim
    •  & Chad M Rienstra
  2. Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

    • Gemma Comellas
    • , Alexander M Barclay
    •  & Chad M Rienstra
  3. Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.

    • Dustin J Covell
    •  & Virginia M Y Lee
  4. Institute on Aging, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.

    • Dustin J Covell
    •  & Virginia M Y Lee
  5. Center for Neurodegenerative Disease Research, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA.

    • Dustin J Covell
    •  & Virginia M Y Lee
  6. Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee, USA.

    • Amy Kendall
    • , William Wan
    •  & Gerald Stubbs
  7. Center for Structural Biology, Vanderbilt University, Nashville, Tennessee, USA.

    • Amy Kendall
    • , William Wan
    •  & Gerald Stubbs
  8. Division of Computational Bioscience, Center for Information Technology, National Institutes of Health, Bethesda, Maryland, USA.

    • Charles D Schwieters
  9. Department of Biological and Experimental Psychology, School of Biological and Chemical Sciences, Queen Mary University of London, London, UK.

    • Julia M George
  10. Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.

    • Chad M Rienstra


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M.D.T. analyzed and collected SSNMR data, performed structure calculations, analyzed structural features and was the primary author on the manuscript. G.C. analyzed and collected SSNMR data, performed MPL data analysis and provided input on the manuscript. A.J.N. analyzed and collected SSNMR data, performed initial structure calculations and provided input on the manuscript. D.J.C. performed the immunofluorescence, biochemical and biophysical assays and helped prepare the manuscript. D.A.B. prepared isotopically labeled samples and provided input on the manuscript. K.D.K. aided in sample preparation and manuscript preparation. J.M.C. created scripts for data conversion, performed analysis of final structures and provided input on the manuscript. J.K.K. contributed to the sample preparation methods. A.M.B. contributed to the mass spectrometry and solution NMR data and analysis. A.K. and W.W. prepared fiber diffraction samples and collected and analyzed fiber diffraction data. G.S. analyzed fiber diffraction data and aided in manuscript preparation. C.D.S. supported the development of structure calculations in XPLOR-NIH. V.M.Y.L., J.M.G. and C.M.R. were the primary investigators, designed the experiments and aided in manuscript preparation, data collection and interpretation.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Chad M Rienstra.

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