Application of a yeast two-hybrid based screening system in the identification of amyloid-beta aggregation inhibitors in pharmaceutical plants

doi:10.5246/jcps.2011.05.065

Abstract

Abstract:

The aggregation of amyloid-beta (Aβ) peptide, has been demonstrated to be critical for the development of Alzheimer's disease (AD). Aβ aggregation inhibitors are thus considered to be drug candidates for AD therapy. In the present study, we developed a novel screening tool based on the yeast two-hybrid system to screen Aβ aggregation inhibitors. The human Aβ42 peptide cDNA was cloned using assembly PCR and inserted into each of the yeast expression plasmids containing either the GAL4 activation domain (GAL4AD) or the DNA-binding domain (GAL4BD). Co-transformation of the above plasmids led to the expression of the fusion proteins GAL4AD-Aβ42 and GAL4BD-Aβ42 in the AH109 yeast strain. The self interaction of Aβ42 fragments reconstructed the GAL4 transcriptor and thus activated the GAL4 responsive transcription of four reporter genes including HIS3, ADE2, lacZ and MEL1. The expression of the reporter genes rendered the multiple auxotrophic yeast cells capable of growing on the synthetic SD media lacking adenine and histidine. Growth arrest was used as a marker for screening Aβ aggregation inhibitors in this system, and the evaluation of Rhodiola species revealed potential resources for the development of Aβ aggregation inhibitors.

Key words: Alzheimer's disease, Aβ, Yeast two-hybrid, Inhibitors, Drug screening

CLC Number:

Supporting:

Li-Wei Wang, Yan-Fang Yang, Ying-Tao Zhang^* . Application of a yeast two-hybrid based screening system in the identification of amyloid-beta aggregation inhibitors in pharmaceutical plants [J]. , DOI: 10.5246/jcps.2011.05.065.

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