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  • The EMBO Journal (2004) 23, 2039 - 2046
  • doi:10.1038/sj.emboj.7600211

Published online: 22 April 2004

NMR of alpha-synuclein–polyamine complexes elucidates the mechanism and kinetics of induced aggregation

Claudio O Fernández1, Wolfgang Hoyer2, Markus Zweckstetter3, Elizabeth A Jares-Erijman4, Vinod Subramaniam5, Christian Griesinger3 and Thomas M Jovin2

  1. LANAIS RMN 300, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina
  2. Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
  3. Department of NMR-based Structural Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
  4. Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
  5. Biophysical Engineering Group, Faculty of Science and Technology, University of Twente, Enschede, The Netherlands

Correspondence to:

Claudio O Fernández, Laboratorio Nacional de Resonancia Magnética, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina. Tel.: +54 11 496 48 253; Fax: +54 11 496 48 253; E-mail: cfernand@ffyb.uba.ar

Thomas M Jovin, Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany. Tel.: + 49 551 201 1382; Fax +49 551 201 1467; E-mail: tjovin@gwdg.de

Received 28 October 2003; Accepted 22 March 2004

The aggregation of alpha-synuclein is characteristic of Parkinson's disease (PD) and other neurodegenerative synucleinopathies. The 140-aa protein is natively unstructured; thus, ligands binding to the monomeric form are of therapeutic interest. Biogenic polyamines promote the aggregation of alpha-synuclein and may constitute endogenous agents modulating the pathogenesis of PD. We characterized the complexes of natural and synthetic polyamines with alpha-synuclein by NMR and assigned the binding site to C-terminal residues 109–140. Dissociation constants were derived from chemical shift perturbations. Greater polyamine charge (+2 right arrow +5) correlated with increased affinity and enhancement of fibrillation, for which we propose a simple kinetic mechanism involving a dimeric nucleation center. According to the analysis, polyamines increase the extent of nucleation by approx104 and the rate of monomer addition approx40-fold. Significant secondary structure is not induced in monomeric alpha-synuclein by polyamines at 15°C. Instead, NMR reveals changes in a region (aa 22–93) far removed from the polyamine binding site and presumed to adopt the beta-sheet conformation characteristic of fibrillar alpha-synuclein. We conclude that the C-terminal domain acts as a regulator of alpha-synuclein aggregation.

  • Keywords:

    • amyloid,
    • fibrillation,
    • Parkinson's disease,
    • spermine