Reversible heat-induced dissociation of β2-microglobulin amyloid fibrils

J. Kardos, András Micsonai, Henriett Pál-Gábor, Éva Petrik, L. Gráf, J. Kovács, Young Ho Lee, Hironobu Naiki, Yuji Goto

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Recent progress in the field of amyloid research indicates that the classical view of amyloid fibrils, being irreversibly formed highly stable structures resistant to perturbating conditions and proteolytic digestion, is getting more complex. We studied the thermal stability and heat-induced depolymerization of amyloid fibrils of β2-microglobulin (β2m), a protein responsible for dialysis-related amyloidosis. We found that freshly polymerized β2m fibrils at 0.1-0.3 mg/mL concentration completely dissociated to monomers upon 10 min incubation at 99 C. Fibril depolymerization was followed by thioflavin-T fluorescence and circular dichroism spectroscopy at various temperatures. Dissociation of β2m fibrils was found to be a reversible and dynamic process reaching equilibrium between fibrils and monomers within minutes. Repolymerization experiments revealed that the number of extendable fibril ends increased significantly upon incubation at elevated temperatures suggesting that the mechanism of fibril unfolding involves two distinct processes: (1) dissociation of monomers from the fibril ends and (2) the breakage of fibrils. The breakage of fibrils may be an important in vivo factor multiplying the number of fibril nuclei and thus affecting the onset and progress of disease. We investigated the effects of some additives and different factors on the stability of amyloid fibrils. Sample aging increased the thermal stability of β2m fibril solution. 0.5 mM SDS completely prevented β2m fibrils from dissociation up to the applied highest temperature of 99 C. The generality of our findings was proved on fibrils of K3 peptide and α-synuclein. Our simple method may also be beneficial for screening and developing amyloid-active compounds for therapeutic purposes.

Original languageEnglish
Pages (from-to)3211-3220
Number of pages10
JournalBiochemistry
Volume50
Issue number15
DOIs
Publication statusPublished - Apr 19 2011

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Amyloid
Hot Temperature
Depolymerization
Monomers
Temperature
Thermodynamic stability
Synucleins
Circular dichroism spectroscopy
Dialysis
Amyloidosis
Circular Dichroism
Digestion
Spectrum Analysis
Screening
Aging of materials
Fluorescence
Peptides
Research
Proteins
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Reversible heat-induced dissociation of β2-microglobulin amyloid fibrils. / Kardos, J.; Micsonai, András; Pál-Gábor, Henriett; Petrik, Éva; Gráf, L.; Kovács, J.; Lee, Young Ho; Naiki, Hironobu; Goto, Yuji.

In: Biochemistry, Vol. 50, No. 15, 19.04.2011, p. 3211-3220.

Research output: Contribution to journalArticle

Kardos, J, Micsonai, A, Pál-Gábor, H, Petrik, É, Gráf, L, Kovács, J, Lee, YH, Naiki, H & Goto, Y 2011, 'Reversible heat-induced dissociation of β2-microglobulin amyloid fibrils', Biochemistry, vol. 50, no. 15, pp. 3211-3220. https://doi.org/10.1021/bi2000017
Kardos, J. ; Micsonai, András ; Pál-Gábor, Henriett ; Petrik, Éva ; Gráf, L. ; Kovács, J. ; Lee, Young Ho ; Naiki, Hironobu ; Goto, Yuji. / Reversible heat-induced dissociation of β2-microglobulin amyloid fibrils. In: Biochemistry. 2011 ; Vol. 50, No. 15. pp. 3211-3220.
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