Formation of dopamine-mediated alpha-synuclein-soluble oligomers requires methionine oxidation

Free Radic Biol Med. 2009 May 15;46(10):1328-37. doi: 10.1016/j.freeradbiomed.2009.02.009. Epub 2009 Feb 25.

Abstract

alpha-Synuclein is the major component of the intracellular Lewy body inclusions present in Parkinson disease (PD) neurons. PD involves the loss of dopaminergic neurons in the substantia nigra and the subsequent depletion of dopamine (DA) in the striatum. DA can inhibit alpha-synuclein fibrillization in vitro and promote alpha-synuclein aggregation into soluble oligomers. We have studied the mechanism by which DA mediates alpha-synuclein aggregation into soluble oligomers. Reacting alpha-synuclein with DA increased the mass of alpha-synuclein by 64 Da. NMR showed that all four methionine residues were oxidized by DA, consistent with the addition of 64 Da. Substituting all four methionines to alanine significantly reduced the formation of DA-mediated soluble oligomers. The (125)YEMPS(129) motif in alpha-synuclein can modulate DA inhibition of alpha-synuclein fibrillization. However, alpha-synuclein ending before the (125)YEMPS(129) motif (residues 1-124) could still form soluble oligomers. The addition of exogenous synthetic YEMPS peptide inhibited the formation of soluble oligomers and resulted in the YEMPS peptide being oxidized. Therefore, the (125)YEMPS(129) acts as an antioxidant rather than interacting directly with DA. Our study defines methionine oxidation as the dominant mechanism by which DA generates soluble alpha-synuclein oligomers and highlights the potential role for oxidative stress in modulating alpha-synuclein aggregation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs / genetics
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Dopamine / metabolism*
  • Humans
  • Lewy Bodies / metabolism
  • Methionine / metabolism
  • Mutagenesis, Site-Directed
  • Neurons / metabolism*
  • Neurons / ultrastructure
  • Oxidation-Reduction
  • Oxidative Stress
  • Parkinson Disease / pathology
  • Parkinson Disease / physiopathology*
  • Peptides / genetics
  • Peptides / metabolism
  • Protein Binding
  • Protein Multimerization*
  • Solubility
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • alpha-Synuclein / genetics
  • alpha-Synuclein / metabolism*

Substances

  • Peptides
  • alpha-Synuclein
  • Methionine
  • Dopamine