Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity

Li, Shaoliang; Shimono, Chisei; Norioka, Naoko; Nakano, Itsuko; Okubo, Tetsuo; Yagi, Yoshiko; Hayashi, Maria; Sato, Yuya; Fujisaki, Hitomi; Hattori, Shunji; Sugiura, Nobuo; Kimata, Koji; Sekiguchi, Kiyotoshi

doi:https://doi.org/10.1074/jbc.M110.177865

Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity

Li, Shaoliang; Shimono, Chisei; Norioka, Naoko et al.

Journal of Biological Chemistry, 2010, 285(47), 36645-36655

Number of Access:879（2025-08-18 14:07 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71421

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Title	Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity
Creator	NRID 1000040252720 1000040252720 CreatorName Li, Shaoliang
	CreatorName Shimono, Chisei
	CreatorName Norioka, Naoko
	CreatorName Nakano, Itsuko
	CreatorName Okubo, Tetsuo
	CreatorName Yagi, Yoshiko
	CreatorName Hayashi, Maria
	CreatorName Sato, Yuya
	CreatorName Fujisaki, Hitomi
	CreatorName Hattori, Shunji
	NRID 1000090454420 1000090454420 CreatorName Sugiura, Nobuo
	NRID 1000010022641 1000010022641 CreatorName Kimata, Koji
	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. Shaoliang Li, Chisei Shimono, Naoko Norioka, Itsuko Nakano, Tetsuo Okubo, Yoshiko Yagi, Maria Hayashi, Yuya Sato, Hitomi Fujisaki, Shunji Hattori, Nobuo Sugiura, Koji Kimata and Kiyotoshi Sekiguchi. Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity. J. Biol. Chem. 2010; 285: 36645-36655 © the American Society for Biochemistry and Molecular Biology
Abstract	Activin A, a member of the transforming growth factor-β family, plays important roles in hormonal homeostasis and embryogenesis. In this study, we produced recombinant human activin A and examined its abilities to bind to extracellular matrix proteins. Recombinant activin A expressed in 293-F cells was purified as complexes of mature dimeric activin A with its pro-region. Among a panel of extracellular matrix proteins tested, recombinant activin A bound to perlecan and agrin, but not to laminins, nidogens, collagens I and IV, fibronectin, and nephronectin. The binding of recombinant activin A to perlecan was inhibited by heparin and high concentrations of NaCl and abolished by heparitinase treatment of perlecan, suggesting that activin A binds to the heparan sulfate chains of perlecan. In support of this possibility, recombinant activin A was capable of directly binding to heparin and heparan sulfate chains. Site-directed mutagenesis of recombinant activin A revealed that clusters of basic amino acid residues, Lys259-Lys263 and Lys270-Lys272, in the pro-region were required for binding to perlecan. Interestingly, deletion of the peptide segment Lys259-Gly277 containing both basic amino acid clusters from the pro-region did not impair the activity of activin A to stimulate Smad-dependent gene expressions, although it completely ablated the perlecan-binding activity. The binding of activin A to basement membrane heparan sulfate proteoglycans through the basic residues in the pro-region was further confirmed by in situ activin A overlay assays using frozen tissue sections. Taken together, the present results indicate that activin A binds to heparan sulfate proteoglycans through its pro-region and thereby regulates its localization within tissues.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	285(47)
Page	36645-36655
Date of Issued	2010-11
Language	English
Handle URL	https://hdl.handle.net/11094/71421
eISSN	1083351X
NCID	AA1202441X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.M110.177865
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. 2010, 285(47), p. 36645-36655
DC.title	Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity
DC.creator	Li, Shaoliang
	Shimono, Chisei
	Norioka, Naoko
	Nakano, Itsuko
	Okubo, Tetsuo
	Yagi, Yoshiko
	Hayashi, Maria
	Sato, Yuya
	Fujisaki, Hitomi
	Hattori, Shunji
	Sugiura, Nobuo
	Kimata, Koji
	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	2010-11
DC.identifier	https://doi.org/10.1074/jbc.M110.177865
DC.description	This research was originally published in the Journal of Biological Chemistry. Shaoliang Li, Chisei Shimono, Naoko Norioka, Itsuko Nakano, Tetsuo Okubo, Yoshiko Yagi, Maria Hayashi, Yuya Sato, Hitomi Fujisaki, Shunji Hattori, Nobuo Sugiura, Koji Kimata and Kiyotoshi Sekiguchi. Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity. J. Biol. Chem. 2010; 285: 36645-36655 © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	Activin A, a member of the transforming growth factor-β family, plays important roles in hormonal homeostasis and embryogenesis. In this study, we produced recombinant human activin A and examined its abilities to bind to extracellular matrix proteins. Recombinant activin A expressed in 293-F cells was purified as complexes of mature dimeric activin A with its pro-region. Among a panel of extracellular matrix proteins tested, recombinant activin A bound to perlecan and agrin, but not to laminins, nidogens, collagens I and IV, fibronectin, and nephronectin. The binding of recombinant activin A to perlecan was inhibited by heparin and high concentrations of NaCl and abolished by heparitinase treatment of perlecan, suggesting that activin A binds to the heparan sulfate chains of perlecan. In support of this possibility, recombinant activin A was capable of directly binding to heparin and heparan sulfate chains. Site-directed mutagenesis of recombinant activin A revealed that clusters of basic amino acid residues, Lys259-Lys263 and Lys270-Lys272, in the pro-region were required for binding to perlecan. Interestingly, deletion of the peptide segment Lys259-Gly277 containing both basic amino acid clusters from the pro-region did not impair the activity of activin A to stimulate Smad-dependent gene expressions, although it completely ablated the perlecan-binding activity. The binding of activin A to basement membrane heparan sulfate proteoglycans through the basic residues in the pro-region was further confirmed by in situ activin A overlay assays using frozen tissue sections. Taken together, the present results indicate that activin A binds to heparan sulfate proteoglycans through its pro-region and thereby regulates its localization within tissues.
DC.format	71421
citation_title	Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity
citation_author	Li, Shaoliang
	Shimono, Chisei
	Norioka, Naoko
	Nakano, Itsuko
	Okubo, Tetsuo
	Yagi, Yoshiko
	Hayashi, Maria
	Sato, Yuya
	Fujisaki, Hitomi
	Hattori, Shunji
	Sugiura, Nobuo
	Kimata, Koji
	Sekiguchi, Kiyotoshi
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	2010-11
citation_journal_title	Journal of Biological Chemistry
citation_volume	285
citation_issue	47
citation_firstpage	36645
citation_lastpage	36655
citation_doi	https://doi.org/10.1074/jbc.M110.177865
citation_public_url	https://hdl.handle.net/11094/71421

Activin A Binds to Perlecan through Its Pro-region That Has Heparin/Heparan Sulfate Binding Activity

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