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Amyloid-β forms fibrils by nucleated conformational conversion of oligomers

Nature Chemical Biology volume 7, pages 602–609 (2011)Cite this article

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Abstract

Amyloid-β amyloidogenesis is reported to occur via a nucleated polymerization mechanism. If this is true, the energetically unfavorable oligomeric nucleus should be very hard to detect. However, many laboratories have detected early nonfibrillar amyloid-β oligomers without observing amyloid fibrils, suggesting that a mechanistic revision may be needed. Here we introduce Cys-Cys-amyloid-β1–40, which cannot bind to the latent fluorophore FlAsH as a monomer, but can bind FlAsH as an nonfibrillar oligomer or as a fibril, rendering the conjugates fluorescent. Through FlAsH monitoring of Cys-Cys-amyloid-β1–40 aggregation, we found that amyloid-β1–40 rapidly and efficiently forms spherical oligomers in vitro (85% yield) that are kinetically competent to slowly convert to amyloid fibrils by a nucleated conformational conversion mechanism. This methodology was used to show that plasmalogen ethanolamine vesicles eliminate the proteotoxicity-associated oligomerization phase of amyloid-β amyloidogenesis while allowing fibril formation, rationalizing how low concentrations of plasmalogen ethanolamine in the brain are epidemiologically linked to Alzheimer's disease.

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Figure 1: Detection of Cys-Cys-amyloid-β1–40 fibrils using FlAsH-EDT2.
Figure 2: Monitoring Aβ1–40 aggregation by FlAsH, ThT and ThT-to-FlAsH fluorescence resonance energy transfer (FRET).
Figure 3: CC-Aβ1–40 aggregates by a nucleated conformational conversion mechanism.
Figure 4: Familial Alzheimer's disease mutant Aβ1–40 aggregation time courses.
Figure 5: Effect of LUVs comprising various lipids on Aβ aggregation.
Figure 6: Free-energy diagram for an Aβ1–40 aggregation reaction proceeding by a nucleated conformational conversion mechanism.

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Acknowledgements

We thank the US National Institutes of Health (NS050636), The Skaggs Institute for Chemical Biology and the Lita Annenberg Hazen Foundation for financial support and C. Fearns for critical feedback on the manuscript.

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  1. Department of Chemistry, The Scripps Research Institute and The Skaggs Institute for Chemical Biology, La Jolla, California, USA

    Jiyong Lee, Elizabeth K Culyba, Evan T Powers & Jeffery W Kelly

  2. Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA

    Jiyong Lee, Elizabeth K Culyba & Jeffery W Kelly

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J.L. designed and carried out experiments, analyzed data and wrote the paper. E.K.C. carried out experiments and wrote the paper. E.T.P. and J.W.K. designed experiments, analyzed data and wrote the paper.

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Correspondence to Jeffery W Kelly.

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J.W.K. is a cofounder, paid consultant and a shareholder of FoldRx Pharmaceuticals (Pfizer) and Proteostasis Therapeutics.

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Lee, J., Culyba, E., Powers, E. et al. Amyloid-β forms fibrils by nucleated conformational conversion of oligomers. Nat Chem Biol 7, 602–609 (2011). https://doi.org/10.1038/nchembio.624

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