Amyloid-β protofibrils differ from amyloid-β aggregates induced in dilute hexafluoroisopropanol in stability and morphology

Michael R. Nichols, Melissa A. Moss, Dana Kim Reed, Stephanie Cratic-McDaniel, Jan H. Hoh, Terrone L. Rosenberry

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

The brains of Alzheimer's disease (AD) patients contain large numbers of amyloid plaques that are rich in fibrils composed of 40- and 42-residue amyloid-β (Aβ) peptides. Several lines of evidence indicate that fibrillar Aβ and especially soluble Aβ aggregates are important in the etiology of AD. Recent reports also stress that amyloid aggregates are polymorphic and that a single polypeptide can fold into multiple amyloid conformations. Here we demonstrate that Aβ-(1-40) can form soluble aggregates with predominant β-structures that differ in stability and morphology. One class of aggregates involved soluble Aβ protofibrils, prepared by vigorous overnight agitation of monomeric Aβ-(1-40) at low ionic strength. Dilution of these aggregation reactions induced disaggregation to monomers as measured by size exclusion chromatography. Protofibril concentrations monitored by thioflavin T fluorescence decreased in at least two kinetic phases, with initial disaggregation (rate constant ∼1 h -1) followed by a much slower secondary phase. Incubation of the reactions without agitation resulted in less disaggregation at slower rates, indicating that the protofibrils became progressively more stable over time. In fact, protofibrils isolated by size exclusion chromatography were completely stable and gave no disaggregation. A second class of soluble Aβ aggregates was generated rapidly (<10 min) in buffered 2% hexafluoroisopropanol (HFIP). These aggregates showed increased thioflavin T fluorescence and were rich in β-structure by circular dichroism. Electron microscopy and atomic force microscopy revealed initial globular clusters that progressed over several days to soluble fibrous aggregates. When diluted out of HFIP, these aggregates initially were very unstable and disaggregated completely within 2 min. However, their stability increased as they progressed to fibers. Relative to Aβ protofibrils, the HFIP-induced aggregates seeded elongation by Aβ monomer deposition very poorly. The techniques used to distinguish these two classes of soluble Aβ aggregates may be useful in characterizing Aβ aggregates formed in vivo.

Original languageEnglish (US)
Pages (from-to)2471-2480
Number of pages10
JournalJournal of Biological Chemistry
Volume280
Issue number4
DOIs
StatePublished - Jan 28 2005

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Amyloid
Gel Chromatography
Alzheimer Disease
Fluorescence
Peptides
Atomic Force Microscopy
Amyloid Plaques
Brain Diseases
Circular Dichroism
Osmolar Concentration
Electron Microscopy
Size exclusion chromatography
hexafluoroisopropanol
Monomers
thioflavin T
Ionic strength
Electron microscopy
Dilution
Conformations
Elongation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Amyloid-β protofibrils differ from amyloid-β aggregates induced in dilute hexafluoroisopropanol in stability and morphology. / Nichols, Michael R.; Moss, Melissa A.; Reed, Dana Kim; Cratic-McDaniel, Stephanie; Hoh, Jan H.; Rosenberry, Terrone L.

In: Journal of Biological Chemistry, Vol. 280, No. 4, 28.01.2005, p. 2471-2480.

Research output: Contribution to journalArticle

Nichols, Michael R. ; Moss, Melissa A. ; Reed, Dana Kim ; Cratic-McDaniel, Stephanie ; Hoh, Jan H. ; Rosenberry, Terrone L. / Amyloid-β protofibrils differ from amyloid-β aggregates induced in dilute hexafluoroisopropanol in stability and morphology. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 4. pp. 2471-2480.
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