Molecular Basis of the Recognition of Nephronectin by Integrin α8β1

Sato, Yuya; Uemura, Toshihiko; Morimitsu, Keisuke; Sato, Ryoko; Manabe, Ri-ichiroh; Takagi, Junichi; Yamada, Masashi; Sekiguchi, Kiyotoshi

doi:https://doi.org/10.1074/jbc.M900200200

Molecular Basis of the Recognition of Nephronectin by Integrin α8β1

Sato, Yuya; Uemura, Toshihiko; Morimitsu, Keisuke et al.

Journal of Biological Chemistry, 2009, 284(21), 14524-14536

Number of Access:980（2025-08-15 06:46 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71424

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Title	Molecular Basis of the Recognition of Nephronectin by Integrin α8β1
Creator	CreatorName Sato, Yuya
	CreatorName Uemura, Toshihiko
	CreatorName Morimitsu, Keisuke
	CreatorName Sato, Ryoko Creator.Alternative Nishiuchi, Ryoko
	NRID 1000030280837 1000030280837 CreatorName Manabe, Ri-ichiroh
	NRID 1000090212000 1000090212000 researchmap permalink read0105692 read0105692 Scopus Author ID 7102173538 7102173538 CreatorName Takagi, Junichi
	NRID 1000090304055 1000090304055 CreatorName Yamada, Masashi
	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. Yuya Sato, Toshihiko Uemura, Keisuke Morimitsu, Ryoko Sato-Nishiuchi, Ri-ichiroh Manabe, Junichi Takagi, Masashi Yamada and Kiyotoshi Sekiguchi. Molecular Basis of the Recognition of Nephronectin by Integrin α8β1. J. Biol. Chem. 2009; 284: 14524-14536 © the American Society for Biochemistry and Molecular Biology
Abstract	Integrin α8β1 interacts with a variety of Arg-Gly-Asp (RGD)-containing ligands in the extracellular matrix. Here, we examined the binding activities of α8β1 integrin toward a panel of RGD-containing ligands. Integrin α8β1 bound specifically to nephronectin with an apparent dissociation constant of 0.28 ± 0.01 nm, but showed only marginal affinities for fibronectin and other RGD-containing ligands. The high-affinity binding to α8β1 integrin was fully reproduced with a recombinant nephronectin fragment derived from the RGD-containing central “linker” segment. A series of deletion mutants of the recombinant fragment identified the LFEIFEIER sequence on the C-terminal side of the RGD motif as an auxiliary site required for high-affinity binding to α8β1 integrin. Alanine scanning mutagenesis within the LFEIFEIER sequence defined the EIE sequence as a critical motif ensuring the high-affinity integrin-ligand interaction. Although a synthetic LFEIFEIER peptide failed to inhibit the binding of α8β1 integrin to nephronectin, a longer peptide containing both the RGD motif and the LFEIFEIER sequence was strongly inhibitory, and was ∼2,000-fold more potent than a peptide containing only the RGD motif. Furthermore, trans-complementation assays using recombinant fragments containing either the RGD motif or LFEIFEIER sequence revealed a clear synergism in the binding to α8β1 integrin. Taken together, these results indicate that the specific high-affinity binding of nephronectin to α8β1 integrin is achieved by bipartite interaction of the integrin with the RGD motif and LFEIFEIER sequence, with the latter serving as a synergy site that greatly potentiates the RGD-driven integrin-ligand interaction but has only marginal activity to secure the interaction by itself.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	284(21)
Page	14524-14536
Date of Issued	2009-05
Language	English
Handle URL	https://hdl.handle.net/11094/71424
eISSN	1083351X
NCID	AA1202441X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.M900200200
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. 2009, 284(21), p. 14524-14536
DC.title	Molecular Basis of the Recognition of Nephronectin by Integrin α8β1
DC.creator	Sato, Yuya
	Uemura, Toshihiko
	Morimitsu, Keisuke
	Sato, Ryoko
	Manabe, Ri-ichiroh
	Takagi, Junichi
	Yamada, Masashi
	Sekiguchi, Kiyotoshi
DC.creator	Nishiuchi, Ryoko
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	2009-05
DC.identifier	https://doi.org/10.1074/jbc.M900200200
DC.description	This research was originally published in the Journal of Biological Chemistry. Yuya Sato, Toshihiko Uemura, Keisuke Morimitsu, Ryoko Sato-Nishiuchi, Ri-ichiroh Manabe, Junichi Takagi, Masashi Yamada and Kiyotoshi Sekiguchi. Molecular Basis of the Recognition of Nephronectin by Integrin α8β1. J. Biol. Chem. 2009; 284: 14524-14536 © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	Integrin α8β1 interacts with a variety of Arg-Gly-Asp (RGD)-containing ligands in the extracellular matrix. Here, we examined the binding activities of α8β1 integrin toward a panel of RGD-containing ligands. Integrin α8β1 bound specifically to nephronectin with an apparent dissociation constant of 0.28 ± 0.01 nm, but showed only marginal affinities for fibronectin and other RGD-containing ligands. The high-affinity binding to α8β1 integrin was fully reproduced with a recombinant nephronectin fragment derived from the RGD-containing central “linker” segment. A series of deletion mutants of the recombinant fragment identified the LFEIFEIER sequence on the C-terminal side of the RGD motif as an auxiliary site required for high-affinity binding to α8β1 integrin. Alanine scanning mutagenesis within the LFEIFEIER sequence defined the EIE sequence as a critical motif ensuring the high-affinity integrin-ligand interaction. Although a synthetic LFEIFEIER peptide failed to inhibit the binding of α8β1 integrin to nephronectin, a longer peptide containing both the RGD motif and the LFEIFEIER sequence was strongly inhibitory, and was ∼2,000-fold more potent than a peptide containing only the RGD motif. Furthermore, trans-complementation assays using recombinant fragments containing either the RGD motif or LFEIFEIER sequence revealed a clear synergism in the binding to α8β1 integrin. Taken together, these results indicate that the specific high-affinity binding of nephronectin to α8β1 integrin is achieved by bipartite interaction of the integrin with the RGD motif and LFEIFEIER sequence, with the latter serving as a synergy site that greatly potentiates the RGD-driven integrin-ligand interaction but has only marginal activity to secure the interaction by itself.
DC.format	71424
citation_title	Molecular Basis of the Recognition of Nephronectin by Integrin α8β1
citation_author	Sato, Yuya
	Uemura, Toshihiko
	Morimitsu, Keisuke
	Sato, Ryoko
	Manabe, Ri-ichiroh
	Takagi, Junichi
	Yamada, Masashi
	Sekiguchi, Kiyotoshi
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	2009-05
citation_journal_title	Journal of Biological Chemistry
citation_volume	284
citation_issue	21
citation_firstpage	14524
citation_lastpage	14536
citation_doi	https://doi.org/10.1074/jbc.M900200200
citation_public_url	https://hdl.handle.net/11094/71424

Molecular Basis of the Recognition of Nephronectin by Integrin α8β1

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