High-throughput analysis of ultrasonication-forced amyloid fibrillation reveals the mechanism underlying the large fluctuation in the lag time

Umemoto, Ayaka; Yagi, Hisashi; So, Masatomo et al.

Journal of Biological Chemistry, 2014, 289(39), 27290-27299.

Number of Access:628（2025-08-30 01:43 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71276

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Title
High-throughput analysis of ultrasonication-forced amyloid fibrillation reveals the mechanism underlying the large fluctuation in the lag time
Creator
CreatorName
Umemoto, Ayaka
NRID
1000010432494 1000010432494
CreatorName
Yagi, Hisashi
NRID
1000040746335 1000040746335
CreatorName
So, Masatomo
NRID
1000040153770 1000040153770
researchmap permalink
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CreatorName
Goto, Yuji
Description
This research was originally published in the Journal of Biological Chemistry. Ayaka Umemoto, Hisashi Yagi, Masatomo So and Yuji Goto. High-throughput Analysis of Ultrasonication-forced Amyloid Fibrillation Reveals the Mechanism Underlying the Large Fluctuation in the Lag Time. J. Biol. Chem. 2014; 289, 27290–27299. © the American Society for Biochemistry and Molecular Biology
Abstract
Amyloid fibrils form in supersaturated solutions of precursor proteins by a nucleation and growth mechanism characterized by a lag time. Although the lag time provides a clue to understanding the complexity of nucleation events, its long period and low reproducibility have been obstacles for exact analysis. Ultrasonication is known to effectively break supersaturation and force fibrillation. By constructing a Handai amyloid burst inducer, which combines a water bath-type ultrasonicator and a microplate reader, we examined the ultrasonication-forced fibrillation of several proteins, with a focus on the fluctuation in the lag time. Amyloid fibrillation of hen egg white lysozyme was examined at pH 2.0 in the presence of 1.0–5.0 M guanidine hydrochloride (GdnHCl), in which the dominant species varied from the native to denatured conformations. Although fibrillation occurred at various concentrations of GdnHCl, the lag time varied largely, with a minimum being observed at ∼3.0 m, the concentration at which GdnHCl-dependent denaturation ended. The coefficient of variation of the lag time did not depend significantly on the GdnHCl concentration and was 2-fold larger than that of the ultrasonication-dependent oxidation of iodide, a simple model reaction. These results suggest that the large fluctuation observed in the lag time for amyloid fibrillation originated from a process associated with a common amyloidogenic intermediate, which may have been a relatively compact denatured conformation. We also suggest that the Handai amyloid burst inducer system will be useful for studying the mechanism of crystallization of proteins because proteins form crystals by the same mechanism as amyloid fibrils under supersaturation.
Publisher
American Society for Biochemistry and Molecular Biology
Source Title
Journal of Biological Chemistry
Volume (Issue)
289(39)
Page
27290-27299.
Date of Issued
2014-09
Language
English
Handle URL
https://hdl.handle.net/11094/71276
eISSN
1083351X
NCID
AA1202445X
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.1074/jbc.M114.569814
Access Rights
open access
oaire:version
Version of Record
Category
学術雑誌論文 Journal Article