1. Academic Validation
  2. Intracellular screening of a peptide library to derive a potent peptide inhibitor of α-synuclein aggregation

Intracellular screening of a peptide library to derive a potent peptide inhibitor of α-synuclein aggregation

  • J Biol Chem. 2015 Mar 20;290(12):7426-35. doi: 10.1074/jbc.M114.620484.
Harish Cheruvara 1 Victoria L Allen-Baume 1 Neil M Kad 2 Jody M Mason 3
Affiliations

Affiliations

  • 1 From the School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex CO4 3SQ.
  • 2 the School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, and.
  • 3 the Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, United Kingdom j.mason@bath.ac.uk.
Abstract

Aggregation of α-synuclein (α-syn) into toxic fibrils is a pathogenic hallmark of Parkinson disease (PD). Studies have focused largely on residues 71-82, yet most early-onset mutations are located between residues 46 and 53. A semirationally designed 209,952-member library based entirely on this region was constructed, containing all wild-type residues and changes associated with early-onset PD. Intracellular cell survival screening and growth competition isolated a 10-residue peptide antagonist that potently inhibits α-syn aggregation and associated toxicity at a 1:1 stoichiometry. This was verified using continuous growth measurements and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytotoxicity studies. Atomic force microscopy and circular dichroism on the same samples showed a random-coil structure and no oligomers. A new region of α-syn for inhibitor targeting has been highlighted, together with the approach of using a semirational design and intracellular screening. The Peptides can then be used as candidates for modification in drugs capable of slowing or even preventing the onset of PD.

Keywords

Alpha-synuclein (α-Synuclein); Amyloid; Library Screening; Parkinson Disease; Protein Misfolding; Protein-Fragment Complementation Assay; Protein-Protein Interaction.

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