Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme

Yamada, Takao; Matsushima, Masaaki; Inaka, Koji; Ohkubo, Tadayasu; Uyeda, Atsuko; Maeda, Toshinaga; Titani, Koiti; Sekiguchi, Kiyotoshi; Kikuchi, Masakazu

doi:https://doi.org/10.1016/S0021-9258(18)82239-6

Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme

Yamada, Takao; Matsushima, Masaaki; Inaka, Koji et al.

Journal of Biological Chemistry, 1993, 268(14), 10588-10592

Number of Access:627（2025-06-28 13:50 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/71440

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Title	Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme
Creator	CreatorName Yamada, Takao
	NRID 1000020242266 1000020242266 CreatorName Matsushima, Masaaki
	CreatorName Inaka, Koji
	NRID 1000090272997 1000090272997 researchmap permalink read0191274 read0191274 Scopus Author ID 7201618682 7201618682 CreatorName Ohkubo, Tadayasu
	CreatorName Uyeda, Atsuko
	NRID 1000020250342 1000020250342 CreatorName Maeda, Toshinaga
	NRID 1000060179942 1000060179942 CreatorName Titani, Koiti
	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 1000030411002 1000030411002 researchmap permalink read0094721 read0094721 CreatorName Kikuchi, Masakazu
Description	This research was originally published in the Journal of Biological Chemistry. T Yamada, M Matsushima, K Inaka, T Ohkubo, A Uyeda, T Maeda, K Titani, K Sekiguchi and M Kikuchi. Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme. J. Biol. Chem. 1993; 268: 10588-10592 © the American Society for Biochemistry and Molecular Biology
Abstract	To determine the functional conformation of the Arg-Gly-Asp (RGD) sequence, we have constructed mutant proteins by inserting 4-12 amino acid residues from the RGD region of human fibronectin between Val74 and Asn75 of human lysozyme. RGDS-, GRGDSP-, TGRGDSPA-, VTGRGDSPAS-, and AVTGRGDS-PASS-introduced mutant lysozymes were expressed in yeast, purified, and designated as RGD4, -6, -8, -10, and -12, respectively. Using baby hamster kidney cells, RGD8, RGD10, and RGD12 were shown to possess high cell adhesion activity nearly equal to 10% of human vitronectin activity. RGD4 and RGD6 exhibited somewhat lower cell adhesion activity. The activities of these mutant proteins were inhibited by the addition of either GRGDSP peptide or polyclonal antibody against vitronectin receptor, as was the case for the vitronectin activity. The results suggest that the cell adhesion signals are transduced to cells through the interaction with the vitronectin receptor. The three-dimensional structures of RGD4 and RGD8 were determined at 1.8-A resolution by x-ray crystallography. A model of the inserted region in RGD4 could be built in the electron density map, but the positions of the preceding residues, Ala73-Val74, were uncertain. The inserted region in RGD8 did not demonstrate continuous electron densities. The results suggest that these RGD sequence-containing regions are highly flexible and that such flexibility could allow the conformation of the RGD regions to be induced to fit into the binding pocket of the integrin receptor.
Publisher	American Society for Biochemistry and Molecular Biology
Source Title	Journal of Biological Chemistry
Volume (Issue)	268(14)
Page	10588-10592
Date of Issued	1993-05
Language	English
Handle URL	https://hdl.handle.net/11094/71440
eISSN	1083351X
NCID	AA1202441X
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1016/S0021-9258(18)82239-6
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. 1993, 268(14), p. 10588-10592
DC.title	Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme
DC.creator	Yamada, Takao
	Matsushima, Masaaki
	Inaka, Koji
	Ohkubo, Tadayasu
	Uyeda, Atsuko
	Maeda, Toshinaga
	Titani, Koiti
	Sekiguchi, Kiyotoshi
	Kikuchi, Masakazu
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	1993-05
DC.identifier	https://doi.org/10.1016/S0021-9258(18)82239-6
DC.description	This research was originally published in the Journal of Biological Chemistry. T Yamada, M Matsushima, K Inaka, T Ohkubo, A Uyeda, T Maeda, K Titani, K Sekiguchi and M Kikuchi. Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme. J. Biol. Chem. 1993; 268: 10588-10592 © the American Society for Biochemistry and Molecular Biology
DCTERMS.abstract	To determine the functional conformation of the Arg-Gly-Asp (RGD) sequence, we have constructed mutant proteins by inserting 4-12 amino acid residues from the RGD region of human fibronectin between Val74 and Asn75 of human lysozyme. RGDS-, GRGDSP-, TGRGDSPA-, VTGRGDSPAS-, and AVTGRGDS-PASS-introduced mutant lysozymes were expressed in yeast, purified, and designated as RGD4, -6, -8, -10, and -12, respectively. Using baby hamster kidney cells, RGD8, RGD10, and RGD12 were shown to possess high cell adhesion activity nearly equal to 10% of human vitronectin activity. RGD4 and RGD6 exhibited somewhat lower cell adhesion activity. The activities of these mutant proteins were inhibited by the addition of either GRGDSP peptide or polyclonal antibody against vitronectin receptor, as was the case for the vitronectin activity. The results suggest that the cell adhesion signals are transduced to cells through the interaction with the vitronectin receptor. The three-dimensional structures of RGD4 and RGD8 were determined at 1.8-A resolution by x-ray crystallography. A model of the inserted region in RGD4 could be built in the electron density map, but the positions of the preceding residues, Ala73-Val74, were uncertain. The inserted region in RGD8 did not demonstrate continuous electron densities. The results suggest that these RGD sequence-containing regions are highly flexible and that such flexibility could allow the conformation of the RGD regions to be induced to fit into the binding pocket of the integrin receptor.
DC.format	71440
citation_title	Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme
citation_author	Yamada, Takao
	Matsushima, Masaaki
	Inaka, Koji
	Ohkubo, Tadayasu
	Uyeda, Atsuko
	Maeda, Toshinaga
	Titani, Koiti
	Sekiguchi, Kiyotoshi
	Kikuchi, Masakazu
citation_publisher	American Society for Biochemistry and Molecular Biology
citation_language	English
citation_date	1993-05
citation_journal_title	Journal of Biological Chemistry
citation_volume	268
citation_issue	14
citation_firstpage	10588
citation_lastpage	10592
citation_doi	https://doi.org/10.1016/S0021-9258(18)82239-6
citation_public_url	https://hdl.handle.net/11094/71440

Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme

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