α-Synuclein Misfolding: Single Molecule AFM Force Spectroscopy Study

Junping Yu, Sarka Malkova, Yuri L Lyubchenko

Research output: Contribution to journalArticle

59 Scopus citations

Abstract

Protein misfolding and aggregation are the very first and critical steps in development of various neurodegenerative disorders, including Parkinson's disease, induced by misfolding of α-synuclein. Thus, elucidating properties of proteins in misfolded states and understanding the mechanisms of their assembly into the disease prone aggregates are critical for the development of rational approaches to prevent protein misfolding-mediated pathologies. To accomplish this goal and as a first step to elucidate the mechanism of α-synuclein misfolding, we applied single-molecule force spectroscopy capable of detecting protein misfolding. We immobilized α-synuclein molecules at their C-termini at the atomic force microscope tips and substrate surfaces, and measured the interaction between the proteins by probing the microscope tip at various locations on the surface. Using this approach, we detected α-synuclein misfolded states by enhanced interprotein interaction. We used a dynamics force spectroscopy approach to measure such an important characteristic of dimers of misfolded α-synuclein as their lifetimes. We found that the dimer lifetimes are in the range of seconds and these values are much higher than the characteristics for the dynamics of the protein in monomeric state. These data show that compared to highly dynamic monomeric forms, α-synuclein dimers are much more stable and thus can serve as stable nuclei for the formation of multimeric and aggregated forms of α-synuclein. Importantly, two different lifetimes were observed for the dimers, suggesting that aggregation can follow different pathways that may lead to different aggregated morphologies of α-synuclein.

Original languageEnglish (US)
Pages (from-to)992-1001
Number of pages10
JournalJournal of Molecular Biology
Volume384
Issue number4
DOIs
StatePublished - Dec 26 2008

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Keywords

  • AFM
  • Parkinson's disease
  • force spectroscopy
  • protein aggregation
  • α-synuclein misfolding

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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