Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10

Ido, Hiroyuki; Harada, Kenji; Futaki, Sugiko; Hayashi, Yoshitaka; Nishiuchi, Ryoko; Natsuka, Yuko; Li, Shaoliang; Wada, Yoshinao; Combs, Ariana C.; Ervasti, James M.; Sekiguchi, Kiyotoshi

doi:https://doi.org/10.1074/jbc.M313626200

Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10

Ido, Hiroyuki; Harada, Kenji; Futaki, Sugiko et al.

Journal of Biological Chemistry, 2004, 279(12), 10946-10954

Number of Access:1,220（2025-08-18 14:07 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71431

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Title	Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10
Creator	CreatorName Ido, Hiroyuki
	CreatorName Harada, Kenji
	NRID 1000000403014 1000000403014 CreatorName Futaki, Sugiko
	NRID 1000080420363 1000080420363 CreatorName Hayashi, Yoshitaka
	CreatorName Nishiuchi, Ryoko
	CreatorName Natsuka, Yuko
	NRID 1000040252720 1000040252720 CreatorName Li, Shaoliang
	NRID 1000000250340 1000000250340 CreatorName Wada, Yoshinao
	CreatorName Combs, Ariana C.
	CreatorName Ervasti, James M.
	NRID 1000050187845 1000050187845 researchmap permalink read0054833 read0054833 Scopus Author ID 7201662709 7201662709 ORCID 0000-0002-6433-1410 0000-0002-6433-1410 CreatorName Sekiguchi, Kiyotoshi
Description	This research was originally published in the Journal of Biological Chemistry. Hiroyuki Ido, Kenji Harada, Sugiko Futaki, Yoshitaka Hayashi, Ryoko Nishiuchi, Yuko Natsuka, Shaoliang Li, Yoshinao Wada, Ariana C. Combs, James M. Ervasti and Kiyotoshi Sekiguchi. Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10. J. Biol. Chem. 2004; 279: 10946-10954 © the American Society for Biochemistry and Molecular Biology
Abstract	The adhesive interactions of cells with laminins are mediated by integrins and non-integrin-type receptors such as α-dystroglycan and syndecans. Laminins bind to these receptors at the C-terminal globular domain of their α chains, but the regions recognized by these receptors have not been mapped precisely. In this study, we sought to locate the binding sites of laminin-10 (α5β1γ1) for α3β1 and α6β1 integrins and α-dystroglycan through the production of a series of recombinant laminin-10 proteins with deletions of the LG (laminin G-like) modules within the globular domain. We found that deletion of the LG4–5 modules did not compromise the binding of laminin-10 to α3β1 and α6β1 integrins but completely abrogated its binding to α-dystroglycan. Further deletion up to the LG3 module resulted in loss of its binding to the integrins, underlining the importance of LG3 for integrin binding by laminin-10. When expressed individually as fusion proteins with glutathione S-transferase or the N-terminal 70-kDa region of fibronectin, only LG4 was capable of binding to α-dystroglycan, whereas neither LG3 nor any of the other LG modules retained the ability to bind to the integrins. Site-directed mutagenesis of the LG3 and LG4 modules indicated that Asp-3198 in the LG3 module is involved in the integrin binding by laminin-10, whereas multiple basic amino acid residues in the putative loop regions are involved synergistically in the α-dystroglycan binding by the LG4 module.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	279(12)
Page	10946-10954
Date of Issued	2004-03
Language	English
Handle URL	https://hdl.handle.net/11094/71431
eISSN	1083351X
NCID	AA1202441X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.M313626200
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. 2004, 279(12), p. 10946-10954
DC.title	Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10
DC.creator	Ido, Hiroyuki
	Harada, Kenji
	Futaki, Sugiko
	Hayashi, Yoshitaka
	Nishiuchi, Ryoko
	Natsuka, Yuko
	Li, Shaoliang
	Wada, Yoshinao
	Combs, Ariana C.
	Ervasti, James M.
	Sekiguchi, Kiyotoshi
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	2004-03
DC.identifier	https://doi.org/10.1074/jbc.M313626200
DC.description	This research was originally published in the Journal of Biological Chemistry. Hiroyuki Ido, Kenji Harada, Sugiko Futaki, Yoshitaka Hayashi, Ryoko Nishiuchi, Yuko Natsuka, Shaoliang Li, Yoshinao Wada, Ariana C. Combs, James M. Ervasti and Kiyotoshi Sekiguchi. Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10. J. Biol. Chem. 2004; 279: 10946-10954 © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	The adhesive interactions of cells with laminins are mediated by integrins and non-integrin-type receptors such as α-dystroglycan and syndecans. Laminins bind to these receptors at the C-terminal globular domain of their α chains, but the regions recognized by these receptors have not been mapped precisely. In this study, we sought to locate the binding sites of laminin-10 (α5β1γ1) for α3β1 and α6β1 integrins and α-dystroglycan through the production of a series of recombinant laminin-10 proteins with deletions of the LG (laminin G-like) modules within the globular domain. We found that deletion of the LG4–5 modules did not compromise the binding of laminin-10 to α3β1 and α6β1 integrins but completely abrogated its binding to α-dystroglycan. Further deletion up to the LG3 module resulted in loss of its binding to the integrins, underlining the importance of LG3 for integrin binding by laminin-10. When expressed individually as fusion proteins with glutathione S-transferase or the N-terminal 70-kDa region of fibronectin, only LG4 was capable of binding to α-dystroglycan, whereas neither LG3 nor any of the other LG modules retained the ability to bind to the integrins. Site-directed mutagenesis of the LG3 and LG4 modules indicated that Asp-3198 in the LG3 module is involved in the integrin binding by laminin-10, whereas multiple basic amino acid residues in the putative loop regions are involved synergistically in the α-dystroglycan binding by the LG4 module.
DC.format	71431
citation_title	Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10
citation_author	Ido, Hiroyuki
	Harada, Kenji
	Futaki, Sugiko
	Hayashi, Yoshitaka
	Nishiuchi, Ryoko
	Natsuka, Yuko
	Li, Shaoliang
	Wada, Yoshinao
	Combs, Ariana C.
	Ervasti, James M.
	Sekiguchi, Kiyotoshi
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	2004-03
citation_journal_title	Journal of Biological Chemistry
citation_volume	279
citation_issue	12
citation_firstpage	10946
citation_lastpage	10954
citation_doi	https://doi.org/10.1074/jbc.M313626200
citation_public_url	https://hdl.handle.net/11094/71431

Molecular Dissection of the α-Dystroglycan- and Integrin-binding Sites within the Globular Domain of Human Laminin-10

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