Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis

Kihara, Miho; Chatani, Eri; Iwata, Kentaro; Yamamoto, Kaori; Matsuura, Takanori; Nakagawa, Atsushi; Naiki, Hironobu; Goto, Yuji

doi:https://doi.org/10.1074/jbc.M605358200

Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis

Kihara, Miho; Chatani, Eri; Iwata, Kentaro et al.

Journal of Biological Chemistry, 2006, 281(41), 31061-31069

Number of Access:448（2025-06-17 08:09 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71292

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Title	Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis
Creator	CreatorName Kihara, Miho
	NRID 1000000432493 1000000432493 CreatorName Chatani, Eri
	CreatorName Iwata, Kentaro
	CreatorName Yamamoto, Kaori
	CreatorName Matsuura, Takanori
	NRID 1000020188890 1000020188890 ORCID 0000-0002-1700-7861 0000-0002-1700-7861 researchmap permalink atsushi_nakagawa atsushi_nakagawa Scopus Author ID 7202934909 7202934909 CreatorName Nakagawa, Atsushi
	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. Miho Kihara, Eri Chatani, Kentaro Iwata, Kaori Yamamoto, Takanori Matsuura, Atsushi Nakagawa, Hironobu Naiki and Yuji Goto. Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis. J. Biol. Chem. 2006; 281, 31061–31069. © the American Society for Biochemistry and Molecular Biology
Abstract	β2-Microglobulin (β2-m), a protein responsible for dialysis-related amyloidosis, adopts an immunoglobulin domain fold in its native state. Although β2-m has Trp residues at positions 60 and 95, both are located near the surface of the domain. Hence, β2-m does not have a conserved Trp common to other immunoglobulin domains, which is buried in close proximity to the disulfide bond. To study the structure of amyloid fibrils in relation to their native fold, we prepared a series of Trp mutants. Trp60 and Trp95 were both replaced with Phe, and a single Trp was introduced at various positions. Among various mutants, W39-β2-m, in which a Trp was introduced at the position corresponding to the conserved Trp, exhibited a remarkable quenching of fluorescence in the native state, as observed for other immunoglobulin domains. An x-ray structural analysis revealed that W39-β2-m assumes the native fold with Trp39 located in the vicinity of the disulfide bond. Comparison of the fluorescence spectra of various mutants for the native and fibrillar forms indicated that, while the Trp residues introduced in the middle of the β2-m sequence tend to be buried in the fibrils, those located in the C-terminal region are more exposed. In addition, the fluorescence spectra of fibrils prepared at pH 2.5 and 7.0 revealed a large difference in the fluorescence intensity for W60-β2-m, implying a major structural difference between them.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	281(41)
Page	31061-31069
Date of Issued	2006-10
Language	English
Handle URL	https://hdl.handle.net/11094/71292
eISSN	1083351X
NCID	AA1202461X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.M605358200
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. 2006, 281(41), p. 31061-31069
DC.title	Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis
DC.creator	Kihara, Miho
	Chatani, Eri
	Iwata, Kentaro
	Yamamoto, Kaori
	Matsuura, Takanori
	Nakagawa, Atsushi
	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	2006-10
DC.identifier	https://doi.org/10.1074/jbc.M605358200
DC.description	This research was originally published in the Journal of Biological Chemistry. Miho Kihara, Eri Chatani, Kentaro Iwata, Kaori Yamamoto, Takanori Matsuura, Atsushi Nakagawa, Hironobu Naiki and Yuji Goto. Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis. J. Biol. Chem. 2006; 281, 31061–31069. © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	β2-Microglobulin (β2-m), a protein responsible for dialysis-related amyloidosis, adopts an immunoglobulin domain fold in its native state. Although β2-m has Trp residues at positions 60 and 95, both are located near the surface of the domain. Hence, β2-m does not have a conserved Trp common to other immunoglobulin domains, which is buried in close proximity to the disulfide bond. To study the structure of amyloid fibrils in relation to their native fold, we prepared a series of Trp mutants. Trp60 and Trp95 were both replaced with Phe, and a single Trp was introduced at various positions. Among various mutants, W39-β2-m, in which a Trp was introduced at the position corresponding to the conserved Trp, exhibited a remarkable quenching of fluorescence in the native state, as observed for other immunoglobulin domains. An x-ray structural analysis revealed that W39-β2-m assumes the native fold with Trp39 located in the vicinity of the disulfide bond. Comparison of the fluorescence spectra of various mutants for the native and fibrillar forms indicated that, while the Trp residues introduced in the middle of the β2-m sequence tend to be buried in the fibrils, those located in the C-terminal region are more exposed. In addition, the fluorescence spectra of fibrils prepared at pH 2.5 and 7.0 revealed a large difference in the fluorescence intensity for W60-β2-m, implying a major structural difference between them.
DC.format	71292
citation_title	Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis
citation_author	Kihara, Miho
	Chatani, Eri
	Iwata, Kentaro
	Yamamoto, Kaori
	Matsuura, Takanori
	Nakagawa, Atsushi
	Naiki, Hironobu
	Goto, Yuji
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	2006-10
citation_journal_title	Journal of Biological Chemistry
citation_volume	281
citation_issue	41
citation_firstpage	31061
citation_lastpage	31069
citation_doi	https://doi.org/10.1074/jbc.M605358200
citation_public_url	https://hdl.handle.net/11094/71292

Conformation of Amyloid Fibrils of β2-Microglobulin Probed by Tryptophan Mutagenesis

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