A high-throughput screen for compounds that inhibit aggregation of the Alzheimer's peptide

ACS Chem Biol. 2006 Aug 22;1(7):461-9. doi: 10.1021/cb600135w.

Abstract

Aggregation of the Alzheimer's peptide Abeta produces toxic multimeric species that play a key role in the development of Alzheimer's disease. Compounds that inhibit this aggregation may prove useful as therapeutic agents for the prevention or treatment of Alzheimer's disease. Although aggregation inhibitors may already exist in combinatorial libraries, finding these compounds in a cost-effective high-throughput manner poses an enormous challenge. To meet this challenge, we have developed a novel high-throughput screen capable of isolating inhibitors of Abeta aggregation from large libraries of inactive candidates. The screen uses a fusion of Abeta42 to GFP. In the absence of inhibition, the rapid misfolding and aggregation of Abeta42 causes the entire fusion protein to misfold, thereby preventing fluorescence. Compounds that inhibit Abeta42 aggregation enable GFP to fold into its native structure and be identified by the resulting fluorescent signal. By implementing the screen on a pilot library of triazine derivatives, we have identified several putative inhibitors. One of the selected compounds was studied in detail by a series of biochemical and biophysical methods. These studies confirmed that the selected compound inhibits aggregation of synthetic Abeta42 peptide. The fluorescence-based method described here is rapid and inexpensive and can be used to screen large libraries for inhibitors of Abeta42 aggregation and/or amyloidogenesis.

MeSH terms

  • Alzheimer Disease / drug therapy*
  • Amyloid beta-Peptides / chemistry
  • Biophysics / methods
  • Chemistry, Pharmaceutical / methods
  • Drug Evaluation, Preclinical*
  • Escherichia coli / metabolism
  • Green Fluorescent Proteins / chemistry
  • Humans
  • Models, Chemical
  • Protein Binding
  • Protein Denaturation
  • Protein Folding
  • Recombinant Fusion Proteins / chemistry
  • Spectrometry, Fluorescence / methods
  • Triazines / chemistry

Substances

  • Amyloid beta-Peptides
  • Recombinant Fusion Proteins
  • Triazines
  • Green Fluorescent Proteins