Oligomers, fact or artefact? SDS-PAGE induces dimerization of β-amyloid in human brain samples

Andrew D. Watt, Keyla A. Perez, Alan Rembach, Nicki A. Sherrat, Lin Wai Hung, Timothy Johanssen, Catriona A. McLean, Woan Mei Kok, Craig A. Hutton, Michelle Fodero-Tavoletti, Colin L. Masters, Victor L. Villemagne, Kevin J. Barnham

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68 Citations (Scopus)

Abstract

The formation of low-order oligomers of β-amyloid (Aβ) within the brain is widely believed to be a central component of Alzheimer's disease (AD) pathogenesis. However, despite advances in high-throughput and high-resolution techniques such as xMAP and mass spectrometry (MS), investigations into these oligomeric species have remained reliant on low-resolution Western blots and enzyme-linked immunosorbent assays. The current investigation compared Aβ profiles within human cortical tissue using sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis (PAGE), xMAP and surface enhanced laser desorption/ionization time-of-flight MS and found that whilst there was significant correlation across the techniques regarding levels of monomeric Aβ, only SDS-PAGE was capable of detecting dimeric isoforms of Aβ. The addition of synthetic di-tyrosine cross-linked Aβ1-40Met 35(O) to the AD tissue demonstrated that the MS methodology was capable of observing dimeric Aβ at femto-molar concentrations, with no noticeable effect on monomeric Aβ levels. Focus turned to the association between SDS-PAGE and levels of observable dimeric Aβ within the AD brain tissue. These investigations revealed that increased levels of dimeric Aβ were observed with increasing concentrations of SDS in the sample buffer. This finding was subsequently confirmed using synthetic Aβ1-42 and suggests that SDS was inducing the formation of dimeric Aβ. The findings that SDS promotes Aβ dimerization have significant implications for the putative role of low-order oligomers in AD pathogenesis and draw into question the utility of oligomeric Aβ as a therapeutic target.

Original languageEnglish
Pages (from-to)549-564
Number of pages16
JournalActa Neuropathologica
Volume125
Issue number4
DOIs
Publication statusPublished - Apr 2013
Externally publishedYes

Keywords

  • Alzheimer's disease
  • Amyloid
  • Dimers
  • Mass spectrometry
  • Oligomers
  • Western blot

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