Abstract
SUMMARYRecent reports point to small soluble oligomers, rather than insoluble fibrils, of amyloid β (Aβ), as the primary toxic species in Alzheimer’s disease. Previously, we developed a low-throughput assay in yeast that is capable of detecting small Aβ42 oligomer formation. Specifically, Aβ42 fused to the functional release factor domain of yeast translational termination factor, Sup35p, formed sodium dodecyl sulfate (SDS)-stable low-n oligomers in living yeast, which impaired release factor activity. As a result, the assay for oligomer formation uses yeast growth to indicate restored release factor activity and presumably reduced oligomer formation. We now describe our translation of this assay into a high-throughput screen (HTS) for anti-oligomeric compounds. By doing so, we also identified two presumptive anti-oligomeric compounds from a sub-library of 12,800 drug-like small molecules. Subsequent biochemical analysis confirmed their anti-oligomeric activity, suggesting that this form of HTS is an efficient, sensitive and cost-effective approach to identify new inhibitors of Aβ42 oligomerization.
Highlights
Several aggregated forms of the amyloid b peptide (Ab), which are generated by proteolytic processing of the amyloid precursor protein (APP), in normal brains and cerebrospinal fluid (CSF) are believed to have a crucial role in the development of Alzheimer’s disease (AD) (Hardy and Higgins, 1992; Selkoe, 1991; Younkin, 1995)
Extracellular amyloid plaques and neurofibrillary tangles formed by insoluble fibrils in brains are hallmarks of AD, recent findings suggest that smaller non-fibrillar oligomeric forms of the Ab peptide are a more likely cause of AD
We examined lysates with non-denaturing PAGE and size exclusion chromatography (SEC; supplementary material Fig. S1), in which the Ab-MRF was detected as a complex with 10-15 monomers and monomers and low-n oligomers were not detected
Summary
Several aggregated forms of the amyloid b peptide (Ab), which are generated by proteolytic processing of the amyloid precursor protein (APP), in normal brains and cerebrospinal fluid (CSF) are believed to have a crucial role in the development of Alzheimer’s disease (AD) (Hardy and Higgins, 1992; Selkoe, 1991; Younkin, 1995). Ab dimers in AD brain or CSF have been identified as toxic because they (but not Ab monomers) induce synaptic dysfunction (Klyubin et al, 2008; Walsh et al, 2002a). Oligomer-specific antibodies can reduce the Abinduced toxicity of soluble AD brain extract (Gong et al, 2003; Lambert et al, 2001; Lee et al, 2006).
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
