N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc

Kariya, Yoshinobu; Kato, Rika; Itoh, Satsuki; Fukuda, Tomohiko; Shibukawa, Yukinao; Sanzen, Noriko; Sekiguchi, Kiyotoshi; Wada, Yoshinao; Kawasaki, Nana; Gu, Jianguo

doi:https://doi.org/10.1074/jbc.M804526200

N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc

Kariya, Yoshinobu; Kato, Rika; Itoh, Satsuki et al.

Journal of Biological Chemistry, 2008, 283(48), 33036-33045

Number of Access:867（2025-08-18 12:36 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71425

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Title	N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc
Creator	NRID 1000000458217 1000000458217 CreatorName Kariya, Yoshinobu
	CreatorName Kato, Rika
	CreatorName Itoh, Satsuki
	NRID 1000040433510 1000040433510 CreatorName Fukuda, Tomohiko
	NRID 1000090393264 1000090393264 CreatorName Shibukawa, Yukinao
	CreatorName Sanzen, Noriko
	NRID 1000050187845 1000050187845 researchmap permalink read0054833 read0054833 Scopus Author ID 7201662709 7201662709 ORCID 0000-0002-6433-1410 0000-0002-6433-1410 CreatorName Sekiguchi, Kiyotoshi
	NRID 1000000250340 1000000250340 CreatorName Wada, Yoshinao
	NRID 1000020186167 1000020186167 CreatorName Kawasaki, Nana
	NRID 1000040260369 1000040260369 CreatorName Gu, Jianguo
Description	This research was originally published in the Journal of Biological Chemistry. Yoshinobu Kariya, Rika Kato, Satsuki Itoh, Tomohiko Fukuda, Yukinao Shibukawa, Noriko Sanzen, Kiyotoshi Sekiguchi, Yoshinao Wada, Nana Kawasaki and Jianguo Gu. N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc. J. Biol. Chem. 2008; 283: 33036-33045 © the American Society for Biochemistry and Molecular Biology
Abstract	Laminin-332 (Lm332) is a large heterotrimeric glycoprotein that has been identified as a scattering factor, a regulator of cancer invasion as well as a prominent basement membrane component of the skin. Past studies have identified the functional domains of Lm332 and revealed the relationships between its activities and the processing of its subunits. However, there is little information available concerning the effects of N-glycosylation on Lm332 activities. In some cancer cells, an increase of β1,6-GlcNAc catalyzed by N-acetylglucosaminyltransferase V (GnT-V) is related to the promotion of cancer cell motility. By contrast, bisecting GlcNAc catalyzed by N-acetylglucosaminyltransferase III (GnT-III) suppresses the further processing with branching enzymes, such as GnT-V, and the elongation of N-glycans. To examine the effects of those N-glycosylations to Lm332 on its activities, we purified Lm332s from the conditioned media of GnT-III- and GnT-V-overexpressing MKN45 cells. Lectin blotting and mass spectrometry analyses revealed that N-glycans containing the bisecting GlcNAc and β1,6-GlcNAc structures were strongly expressed on Lm332 purified from GnT-III-overexpressing (GnT-III-Lm332) and GnT-V-overexpressing (GnT-V-Lm332) cells, respectively. Interestingly, the cell adhesion activity of GnT-III-Lm332 was apparently decreased compared with those of control Lm332 and GnT-V-Lm332. In addition, the introduction of bisecting GlcNAc to Lm332 resulted in a decrease in its cell scattering and migration activities. The weakened activities were most likely derived from the impaired α3β1 integrin clustering and resultant focal adhesion formation. Taken together, our results clearly demonstrate for the first time that N-glycosylation may regulate the biological function of Lm332. This finding could introduce a new therapeutic strategy for cancer.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	283(48)
Page	33036-33045
Date of Issued	2008-11
Language	English
Handle URL	https://hdl.handle.net/11094/71425
eISSN	1083351X
NCID	AA1202441X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1074/jbc.M804526200
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. 2008, 283(48), p. 33036-33045
DC.title	N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc
DC.creator	Kariya, Yoshinobu
	Kato, Rika
	Itoh, Satsuki
	Fukuda, Tomohiko
	Shibukawa, Yukinao
	Sanzen, Noriko
	Sekiguchi, Kiyotoshi
	Wada, Yoshinao
	Kawasaki, Nana
	Gu, Jianguo
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	2008-11
DC.identifier	https://doi.org/10.1074/jbc.M804526200
DC.description	This research was originally published in the Journal of Biological Chemistry. Yoshinobu Kariya, Rika Kato, Satsuki Itoh, Tomohiko Fukuda, Yukinao Shibukawa, Noriko Sanzen, Kiyotoshi Sekiguchi, Yoshinao Wada, Nana Kawasaki and Jianguo Gu. N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc. J. Biol. Chem. 2008; 283: 33036-33045 © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	Laminin-332 (Lm332) is a large heterotrimeric glycoprotein that has been identified as a scattering factor, a regulator of cancer invasion as well as a prominent basement membrane component of the skin. Past studies have identified the functional domains of Lm332 and revealed the relationships between its activities and the processing of its subunits. However, there is little information available concerning the effects of N-glycosylation on Lm332 activities. In some cancer cells, an increase of β1,6-GlcNAc catalyzed by N-acetylglucosaminyltransferase V (GnT-V) is related to the promotion of cancer cell motility. By contrast, bisecting GlcNAc catalyzed by N-acetylglucosaminyltransferase III (GnT-III) suppresses the further processing with branching enzymes, such as GnT-V, and the elongation of N-glycans. To examine the effects of those N-glycosylations to Lm332 on its activities, we purified Lm332s from the conditioned media of GnT-III- and GnT-V-overexpressing MKN45 cells. Lectin blotting and mass spectrometry analyses revealed that N-glycans containing the bisecting GlcNAc and β1,6-GlcNAc structures were strongly expressed on Lm332 purified from GnT-III-overexpressing (GnT-III-Lm332) and GnT-V-overexpressing (GnT-V-Lm332) cells, respectively. Interestingly, the cell adhesion activity of GnT-III-Lm332 was apparently decreased compared with those of control Lm332 and GnT-V-Lm332. In addition, the introduction of bisecting GlcNAc to Lm332 resulted in a decrease in its cell scattering and migration activities. The weakened activities were most likely derived from the impaired α3β1 integrin clustering and resultant focal adhesion formation. Taken together, our results clearly demonstrate for the first time that N-glycosylation may regulate the biological function of Lm332. This finding could introduce a new therapeutic strategy for cancer.
DC.format	71425
citation_title	N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc
citation_author	Kariya, Yoshinobu
	Kato, Rika
	Itoh, Satsuki
	Fukuda, Tomohiko
	Shibukawa, Yukinao
	Sanzen, Noriko
	Sekiguchi, Kiyotoshi
	Wada, Yoshinao
	Kawasaki, Nana
	Gu, Jianguo
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	2008-11
citation_journal_title	Journal of Biological Chemistry
citation_volume	283
citation_issue	48
citation_firstpage	33036
citation_lastpage	33045
citation_doi	https://doi.org/10.1074/jbc.M804526200
citation_public_url	https://hdl.handle.net/11094/71425

N-Glycosylation of Laminin-332 Regulates Its Biological Functions : A NOVEL FUNCTION OF THE BISECTING GlcNAc

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