Validation of folate in a convenient yeast assay suited for identification of inhibitors of Alzheimer's amyloid-beta aggregation

J Alzheimers Dis. 2008 Nov;15(3):391-6. doi: 10.3233/jad-2008-15305.

Abstract

Previously in the search for chemopreventatives for Alzheimer's disease (AD), microbial cells with amyloid-beta (Abeta) protein fusions have been used to screen for compounds that reduce the aggregation, misfolding or oligomerization of Abeta. In the current study, such a system has been used to look at the effect of folate, whose deficiency has been associated with AD. A folate-deficient yeast strain producing Abeta fused to green fluorescent protein (GFP) that spontaneously misfolds/aggregates and suppresses green fluorescence was used to examine the effects of folinic acid on Abeta-GFP fluorescence. At levels that did not affect growth or plasmid stability, increasing levels of folinic acid led to increasing green fluorescence, suggesting folate can assist with preventing Abeta-misfolding/aggregation. This result supports a protective role for folate and suggests that yeast assays may provide significant new approaches for testing of AD chemopreventatives.

MeSH terms

  • Alzheimer Disease / genetics*
  • Alzheimer Disease / metabolism*
  • Amyloid beta-Peptides / chemistry*
  • Amyloid beta-Peptides / drug effects*
  • Biological Assay
  • Flow Cytometry
  • Folic Acid / analysis*
  • Gene Deletion
  • Green Fluorescent Proteins / genetics
  • Leucovorin / pharmacology
  • Peptide Synthases / genetics
  • Plasmids / genetics
  • Protein Folding / drug effects
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism*
  • Vitamin B Complex / pharmacology

Substances

  • Amyloid beta-Peptides
  • Vitamin B Complex
  • Green Fluorescent Proteins
  • Folic Acid
  • Peptide Synthases
  • dihydrofolate synthetase
  • Leucovorin