Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence

Ban, Tadato; Hamada, Daizo; Hasegawa, Kazuhiro; Naiki, Hironobu; Goto, Yuji

doi:https://doi.org/10.1074/jbc.C300049200

Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence

Ban, Tadato; Hamada, Daizo; Hasegawa, Kazuhiro et al.

Journal of Biological Chemistry, 2003, 278(19), 16462-16465

Number of Access:606（2025-08-30 01:41 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71300

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Title	Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence
Creator	NRID 1000000579667 1000000579667 CreatorName Ban, Tadato
	NRID 1000060372132 1000060372132 CreatorName Hamada, Daizo
	NRID 1000060324159 1000060324159 CreatorName Hasegawa, Kazuhiro
	NRID 1000010227704 1000010227704 CreatorName Naiki, Hironobu
	NRID 1000040153770 1000040153770 researchmap permalink read0014440 read0014440 CreatorName Goto, Yuji
Description	This research was originally published in the Journal of Biological Chemistry. Tadato Ban, Daizo Hamada, Kazuhiro Hasegawa, Hironobu Naiki and Yuji Goto. Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence. J. Biol. Chem. 2003; 278, 16462-16465. © the American Society for Biochemistry and Molecular Biology
Abstract	Real-time monitoring of fibril growth is essential to clarify the mechanism of amyloid fibril formation. Thioflavin T (ThT) is a reagent known to become strongly fluorescent upon binding to amyloid fibrils. Here, we show that, by monitoring ThT fluorescence with total internal reflection fluorescence microscopy (TIRFM), amyloid fibrils of β2-microgobulin (β2-m) can be visualized without requiring covalent fluorescence labeling. One of the advantages of TIRFM would be that we selectively monitor fibrils lying along the slide glass, so that we can obtain the exact length of fibrils. This method was used to follow the kinetics of seed-dependent β2-m fibril extension. The extension was unidirectional with various rates, suggesting the heterogeneity of the amyloid structures. Since ThT binding is common to all amyloid fibrils, the present method will have general applicability for the analysis of amyloid fibrils. We confirmed this with the octapeptide corresponding to the C terminus derived from human medin and the Alzheimer's amyloid β-peptide.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	278(19)
Page	16462-16465
Date of Issued	2003-05
Language	English
Handle URL	https://hdl.handle.net/11094/71300
eISSN	1083351X
NCID	AA1202469X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.C300049200
Access Rights	open access
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	Journal of Biological Chemistry. 2003, 278(19), p. 16462-16465
DC.title	Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence
DC.creator	Ban, Tadato
	Hamada, Daizo
	Hasegawa, Kazuhiro
	Naiki, Hironobu
	Goto, Yuji
DC.publisher	American Society for Biochemistry and Molecular Biology
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2003-05
DC.identifier	https://doi.org/10.1074/jbc.C300049200
DC.description	This research was originally published in the Journal of Biological Chemistry. Tadato Ban, Daizo Hamada, Kazuhiro Hasegawa, Hironobu Naiki and Yuji Goto. Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence. J. Biol. Chem. 2003; 278, 16462-16465. © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	Real-time monitoring of fibril growth is essential to clarify the mechanism of amyloid fibril formation. Thioflavin T (ThT) is a reagent known to become strongly fluorescent upon binding to amyloid fibrils. Here, we show that, by monitoring ThT fluorescence with total internal reflection fluorescence microscopy (TIRFM), amyloid fibrils of β2-microgobulin (β2-m) can be visualized without requiring covalent fluorescence labeling. One of the advantages of TIRFM would be that we selectively monitor fibrils lying along the slide glass, so that we can obtain the exact length of fibrils. This method was used to follow the kinetics of seed-dependent β2-m fibril extension. The extension was unidirectional with various rates, suggesting the heterogeneity of the amyloid structures. Since ThT binding is common to all amyloid fibrils, the present method will have general applicability for the analysis of amyloid fibrils. We confirmed this with the octapeptide corresponding to the C terminus derived from human medin and the Alzheimer's amyloid β-peptide.
DC.format	71300
citation_title	Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence
citation_author	Ban, Tadato
	Hamada, Daizo
	Hasegawa, Kazuhiro
	Naiki, Hironobu
	Goto, Yuji
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	2003-05
citation_journal_title	Journal of Biological Chemistry
citation_volume	278
citation_issue	19
citation_firstpage	16462
citation_lastpage	16465
citation_doi	https://doi.org/10.1074/jbc.C300049200
citation_public_url	https://hdl.handle.net/11094/71300

Direct Observation of Amyloid Fibril Growth Monitored by Thioflavin T Fluorescence

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