Anti-amyloid compounds inhibit α-synuclein aggregation induced by protein misfolding cyclic amplification (PMCA)

Research output: Contribution to journalArticle

Abstract

Filaments made of α-synuclein form the characteristic Lewy pathology in Parkinson and other diseases. The formation of α-synuclein filaments can be reproduced in vitro by incubation of recombinant protein, but the filament growth is very slow and highly variable and so unsuitable for fast high throughput anti-aggregation drug screening. To overcome this obstacle we have investigated whether the protein misfolding cyclic amplification (PMCA) technique, used for fast amplification of prion protein aggregates, could be adapted for growing α-synuclein aggregates and thus suitable for screening of drugs to affect α-synuclein aggregation for the treatment of the yet incurable α-synucleinopathies. Circular dichroism, electron microscopy, and native and SDS-polyacrylamide gels were used to demonstrate α-synuclein aggregate formation by PMCA, and the strain imprint of the α-synuclein fibrils was studied by proteinase K digestion. We also demonstrated that α-synuclein fibrils are able to seed new α-synuclein PMCA reactions and to enter and aggregate in cells in culture. In particular, we have generated a line of "chronically infected" cells, which transmit α-synuclein aggregates even after multiple passages. To evaluate the sensitivity of the PMCA system as an α-synuclein anti-aggregating drug screening assay a panel of 10 drugs was tested. Anti-amyloid compounds proved efficient in inhibiting α-synuclein fibril formation induced by PMCA. Our results show that α-synuclein PMCA is a fast and reproducible system that could be used as a high throughput screening method for finding new α-synuclein anti-aggregating compounds.

Details

Authors
  • Maria Eugenia Herva
  • Shahin Zibaee
  • Graham Fraser
  • Roger A Barker
  • Michel Goedert
  • Maria Grazia Spillantini
Organisations
External organisations
  • University of Cambridge
Research areas and keywords

Keywords

  • Amyloid, Cell Line, Humans, Protein Folding, Recombinant Proteins, Reproducibility of Results, alpha-Synuclein, Journal Article, Research Support, Non-U.S. Gov't
Original languageEnglish
Pages (from-to)11897-905
Number of pages9
JournalJournal of Biological Chemistry
Volume289
Issue number17
Publication statusPublished - 2014 Apr 25
Publication categoryResearch
Peer-reviewedYes