NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP

Mizushima, Ryota; Kim, Ju Yaen; Suetake, Isao; Tanaka, Hiroaki; Takai, Tomoyo; Kamiya, Narutoshi; Takano, Yu; Mishima, Yuichi; Tajima, Shoji; Goto, Yuji; Fukui, Kenji; Lee, Young-Ho

doi:https://doi.org/10.1371/journal.pone.0098554

NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP

Mizushima, Ryota; Kim, Ju Yaen; Suetake, Isao et al.

PLOS ONE, 2014, 9(6), e98554

Number of Access:1,106（2025-08-30 01:43 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71261

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Title	NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP
Creator	CreatorName Mizushima, Ryota
	CreatorName Kim, Ju Yaen
	NRID 1000080304054 1000080304054 CreatorName Suetake, Isao
	CreatorName Tanaka, Hiroaki
	CreatorName Takai, Tomoyo
	NRID 1000080420462 1000080420462 CreatorName Kamiya, Narutoshi
	NRID 1000030403017 1000030403017 CreatorName Takano, Yu
	CreatorName Mishima, Yuichi
	NRID 1000050132931 1000050132931 CreatorName Tajima, Shoji
	NRID 1000040153770 1000040153770 researchmap permalink read0014440 read0014440 CreatorName Goto, Yuji
	NRID 1000000466038 1000000466038 CreatorName Fukui, Kenji
	NRID 1000070589431 1000070589431 CreatorName Lee, Young-Ho
Description	Mizushima R, Kim JY, Suetake I, Tanaka H, Takai T, et al. (2014) NMR Characterization of the Interaction of the Endonuclease Domain of MutL withDivalent Metal Ions and ATP. PLoS ONE 9(6): e98554. doi:10.1371/journal.pone.0098554
Abstract	MutL is a multi-domain protein comprising an N-terminal ATPase domain (NTD) and C-terminal dimerization domain (CTD), connected with flexible linker regions, that plays a key role in DNA mismatch repair. To expand understanding of the regulation mechanism underlying MutL endonuclease activity, our NMR-based study investigated interactions between the CTD of MutL, derived from the hyperthermophilic bacterium Aquifex aeolicus (aqMutL-CTD), and putative binding molecules. Chemical shift perturbation analysis with the model structure of aqMutL-CTD and circular dichroism results revealed that tight Zn2+ binding increased thermal stability without changing secondary structures to function at high temperatures. Peak intensity analysis exploiting the paramagnetic relaxation enhancement effect indicated the binding site for Mn2+, which shared binding sites for Zn2+. The coexistence of these two metal ions appears to be important for the function of MutL. Chemical shift perturbation analysis revealed a novel ATP binding site in aqMutL-CTD. A docking simulation incorporating the chemical shift perturbation data provided a putative scheme for the intermolecular interactions between aqMutL-CTD and ATP. We proposed a simple and understandable mechanical model for the regulation of MutL endonuclease activity in MMR based on the relative concentrations of ATP and CTD through ATP binding-regulated interdomain interactions between CTD and NTD.
Publisher	PLOS
Source Title	PLOS ONE
Volume (Issue)	9(6)
Page	e98554
Date of Issued	2014-06
Language	English
Handle URL	https://hdl.handle.net/11094/71261
eISSN	19326203
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1371/journal.pone.0098554
Access Rights	open access
CopyRight	© 2014 Mizushima et al. This article is licensed under a Creative Commons Attribution 4.0 International License.
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	PLOS ONE. 2014, 9(6), p. e98554
DC.title	NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP
DC.creator	Mizushima, Ryota
	Kim, Ju Yaen
	Suetake, Isao
	Tanaka, Hiroaki
	Takai, Tomoyo
	Kamiya, Narutoshi
	Takano, Yu
	Mishima, Yuichi
	Tajima, Shoji
	Goto, Yuji
	Fukui, Kenji
	Lee, Young-Ho
DC.publisher	PLOS
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2014-06
DC.identifier	https://doi.org/10.1371/journal.pone.0098554
DC.description	Mizushima R, Kim JY, Suetake I, Tanaka H, Takai T, et al. (2014) NMR Characterization of the Interaction of the Endonuclease Domain of MutL withDivalent Metal Ions and ATP. PLoS ONE 9(6): e98554. doi:10.1371/journal.pone.0098554
DCTERMS.abstract	MutL is a multi-domain protein comprising an N-terminal ATPase domain (NTD) and C-terminal dimerization domain (CTD), connected with flexible linker regions, that plays a key role in DNA mismatch repair. To expand understanding of the regulation mechanism underlying MutL endonuclease activity, our NMR-based study investigated interactions between the CTD of MutL, derived from the hyperthermophilic bacterium Aquifex aeolicus (aqMutL-CTD), and putative binding molecules. Chemical shift perturbation analysis with the model structure of aqMutL-CTD and circular dichroism results revealed that tight Zn2+ binding increased thermal stability without changing secondary structures to function at high temperatures. Peak intensity analysis exploiting the paramagnetic relaxation enhancement effect indicated the binding site for Mn2+, which shared binding sites for Zn2+. The coexistence of these two metal ions appears to be important for the function of MutL. Chemical shift perturbation analysis revealed a novel ATP binding site in aqMutL-CTD. A docking simulation incorporating the chemical shift perturbation data provided a putative scheme for the intermolecular interactions between aqMutL-CTD and ATP. We proposed a simple and understandable mechanical model for the regulation of MutL endonuclease activity in MMR based on the relative concentrations of ATP and CTD through ATP binding-regulated interdomain interactions between CTD and NTD.
DC.rights	© 2014 Mizushima et al. This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	71261
citation_title	NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP
citation_author	Mizushima, Ryota
	Kim, Ju Yaen
	Suetake, Isao
	Tanaka, Hiroaki
	Takai, Tomoyo
	Kamiya, Narutoshi
	Takano, Yu
	Mishima, Yuichi
	Tajima, Shoji
	Goto, Yuji
	Fukui, Kenji
	Lee, Young-Ho
citation_publisher	PLOS
citation_language	English
citation_date	2014-06
citation_journal_title	PLOS ONE
citation_volume	9
citation_issue	6
citation_firstpage	e98554
citation_doi	https://doi.org/10.1371/journal.pone.0098554
citation_public_url	https://hdl.handle.net/11094/71261

NMR Characterization of the Interaction of the Endonuclease Domain of MutL with Divalent Metal Ions and ATP

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