A pH-dependent switch promotes β-synuclein fibril formation via glutamate residues

J Biol Chem. 2017 Sep 29;292(39):16368-16379. doi: 10.1074/jbc.M117.780528. Epub 2017 Jul 14.


α-Synuclein (αS) is the primary protein associated with Parkinson's disease, and it undergoes aggregation from its intrinsically disordered monomeric form to a cross-β fibrillar form. The closely related homolog β-synuclein (βS) is essentially fibril-resistant under cytoplasmic physiological conditions. Toxic gain-of-function by βS has been linked to dysfunction, but the aggregation behavior of βS under altered pH is not well-understood. In this work, we compare fibril formation of αS and βS at pH 7.3 and mildly acidic pH 5.8, and we demonstrate that pH serves as an on/off switch for βS fibrillation. Using αS/βS domain-swapped chimera constructs and single residue substitutions in βS, we localized the switch to acidic residues in the N-terminal and non-amyloid component domains of βS. Computational models of βS fibril structures indicate that key glutamate residues (Glu-31 and Glu-61) in these domains may be sites of pH-sensitive interactions, and variants E31A and E61A show dramatically altered pH sensitivity for fibril formation supporting the importance of these charged side chains in fibril formation of βS. Our results demonstrate that relatively small changes in pH, which occur frequently in the cytoplasm and in secretory pathways, may induce the formation of βS fibrils and suggest a complex role for βS in synuclein cellular homeostasis and Parkinson's disease.

Keywords: Parkinson disease; beta-synuclein; computer modeling; fibril; neurodegeneration; nuclear magnetic resonance (NMR); protein aggregation; synuclein.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Substitution
  • Glutamic Acid / chemistry*
  • Humans
  • Hydrogen Bonding
  • Hydrogen-Ion Concentration
  • Microfibrils / chemistry
  • Microfibrils / metabolism
  • Microfibrils / pathology
  • Models, Molecular*
  • Mutagenesis, Site-Directed
  • Parkinson Disease / metabolism
  • Parkinson Disease / pathology
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Point Mutation
  • Protein Aggregation, Pathological / genetics
  • Protein Aggregation, Pathological / metabolism*
  • Protein Aggregation, Pathological / pathology
  • Protein Interaction Domains and Motifs
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • alpha-Synuclein / chemistry
  • alpha-Synuclein / genetics
  • alpha-Synuclein / metabolism
  • beta-Synuclein / chemistry
  • beta-Synuclein / genetics
  • beta-Synuclein / metabolism*


  • Peptide Fragments
  • Recombinant Fusion Proteins
  • SNCA protein, human
  • SNCB protein, human
  • alpha-Synuclein
  • beta-Synuclein
  • Glutamic Acid

Associated data

  • PDB/2n0a