Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange

Lee, Young-Ho; Tamura, Kosuke; Maeda, Masahiro; Hoshino, Masaru; Sakurai, Kazumasa; Takahashi, Satoshi; Ikegami, Takahisa; Hase, Toshiharu; Goto, Yuji

doi:https://doi.org/10.1074/jbc.M608417200

Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange

Lee, Young-Ho; Tamura, Kosuke; Maeda, Masahiro et al.

Journal of Biological Chemistry, 2007, 282(8), 5959-5967

Number of Access:1,197（2025-08-30 10:43 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71290

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Title	Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange
Creator	NRID 1000070589431 1000070589431 CreatorName Lee, Young-Ho
	CreatorName Tamura, Kosuke
	CreatorName Maeda, Masahiro
	NRID 1000070304053 1000070304053 CreatorName Hoshino, Masaru
	NRID 1000010403015 1000010403015 CreatorName Sakurai, Kazumasa
	NRID 1000030283641 1000030283641 CreatorName Takahashi, Satoshi
	NRID 1000020283939 1000020283939 CreatorName Ikegami, Takahisa
	NRID 1000000127276 1000000127276 CreatorName Hase, Toshiharu
	NRID 1000040153770 1000040153770 researchmap permalink read0014440 read0014440 CreatorName Goto, Yuji
Description	This research was originally published in the Journal of Biological Chemistry. Young-Ho Lee, Kosuke Tamura, Masahiro Maeda, Masaru Hoshino, Kazumasa Sakurai, Satoshi Takahashi, Takahisa Ikegami, Toshiharu Hase, and Yuji Goto. Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange. J. Biol. Chem. 2007; 282, 5959-5967. © the American Society for Biochemistry and Molecular Biology
Abstract	NMR-detected hydrogen/deuterium (H/D) exchange of amide protons is a powerful way for investigating the residue-based conformational stability and dynamics of proteins in solution. Maize ferredoxin-NADP+ reductase (FNR) is a relatively large protein with 314 amino acid residues, consisting of flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide phosphate (NADP+)-binding domains. To address the structural stability and dynamics of FNR, H/D exchange of amide protons was performed using heteronuclear NMR at pDr values 8.0 and 6.0, physiologically relevant conditions mimicking inside of chloroplasts. At both pDr values, the exchange rate varied widely depending on the residues. The profiles of protected residues revealed that the highly protected regions matched well with the hydrophobic cores suggested from the crystal structure, and that the NADP+-binding domain can be divided into two subdomains. The global stability of FNR obtained by H/D exchange with NMR was higher than that by chemical denaturation, indicating that H/D exchange is especially useful for analyzing the residue-based conformational stability of large proteins, for which global unfolding is mostly irreversible. Interestingly, more dynamic conformation of the C-terminal subdomain of the NADP+-binding domain at pDr 8.0, the daytime pH in chloroplasts, than at pDr 6.0 is likely to be involved in the increased binding of NADP+ for elevating the activity of FNR. In light of photosynthesis, the present study provides the first structure-based relationship of dynamics with function for the FNR-type family in solution.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	282(8)
Page	5959-5967
Date of Issued	2007-02
Language	English
Handle URL	https://hdl.handle.net/11094/71290
eISSN	1083351X
NCID	AA1202459X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.M608417200
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. 2007, 282(8), p. 5959-5967
DC.title	Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange
DC.creator	Lee, Young-Ho
	Tamura, Kosuke
	Maeda, Masahiro
	Hoshino, Masaru
	Sakurai, Kazumasa
	Takahashi, Satoshi
	Ikegami, Takahisa
	Hase, Toshiharu
	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	2007-02
DC.identifier	https://doi.org/10.1074/jbc.M608417200
DC.description	This research was originally published in the Journal of Biological Chemistry. Young-Ho Lee, Kosuke Tamura, Masahiro Maeda, Masaru Hoshino, Kazumasa Sakurai, Satoshi Takahashi, Takahisa Ikegami, Toshiharu Hase, and Yuji Goto. Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange. J. Biol. Chem. 2007; 282, 5959-5967. © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	NMR-detected hydrogen/deuterium (H/D) exchange of amide protons is a powerful way for investigating the residue-based conformational stability and dynamics of proteins in solution. Maize ferredoxin-NADP+ reductase (FNR) is a relatively large protein with 314 amino acid residues, consisting of flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide phosphate (NADP+)-binding domains. To address the structural stability and dynamics of FNR, H/D exchange of amide protons was performed using heteronuclear NMR at pDr values 8.0 and 6.0, physiologically relevant conditions mimicking inside of chloroplasts. At both pDr values, the exchange rate varied widely depending on the residues. The profiles of protected residues revealed that the highly protected regions matched well with the hydrophobic cores suggested from the crystal structure, and that the NADP+-binding domain can be divided into two subdomains. The global stability of FNR obtained by H/D exchange with NMR was higher than that by chemical denaturation, indicating that H/D exchange is especially useful for analyzing the residue-based conformational stability of large proteins, for which global unfolding is mostly irreversible. Interestingly, more dynamic conformation of the C-terminal subdomain of the NADP+-binding domain at pDr 8.0, the daytime pH in chloroplasts, than at pDr 6.0 is likely to be involved in the increased binding of NADP+ for elevating the activity of FNR. In light of photosynthesis, the present study provides the first structure-based relationship of dynamics with function for the FNR-type family in solution.
DC.format	71290
citation_title	Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange
citation_author	Lee, Young-Ho
	Tamura, Kosuke
	Maeda, Masahiro
	Hoshino, Masaru
	Sakurai, Kazumasa
	Takahashi, Satoshi
	Ikegami, Takahisa
	Hase, Toshiharu
	Goto, Yuji
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	2007-02
citation_journal_title	Journal of Biological Chemistry
citation_volume	282
citation_issue	8
citation_firstpage	5959
citation_lastpage	5967
citation_doi	https://doi.org/10.1074/jbc.M608417200
citation_public_url	https://hdl.handle.net/11094/71290

Cores and pH-dependent Dynamics of Ferredoxin-NADP+ Reductase Revealed by Hydrogen/Deuterium Exchange

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