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

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タイトル	Structural and functional analyses of the Arg-Gly-Asp sequence introduced into human lysozyme
著者	Yamada, Takao Yamada, Takao
	Matsushima, Masaaki Matsushima, Masaaki
	Inaka, Koji Inaka, Koji
	Ohkubo, Tadayasu Ohkubo, Tadayasu
	Uyeda, Atsuko Uyeda, Atsuko
	Maeda, Toshinaga Maeda, Toshinaga
	Titani, Koiti Titani, Koiti
	Sekiguchi, Kiyotoshi Sekiguchi, Kiyotoshi
	Kikuchi, Masakazu Kikuchi, Masakazu
内容	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
抄録	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.
公開者	American Society for Biochemistry and Molecular Biology
掲載誌名	Journal of Biological Chemistry
巻	268
号	14
開始ページ	10588
終了ページ	10592
刊行年月	1993-05
ISSN	1083351X
NCID	AA1202441X
URL	http://hdl.handle.net/11094/71440
関連情報 (references)	http://www.jbc.org/content/268/14/10588
言語	英語
カテゴリ	学術雑誌論文 Journal Article

著者版フラグ	publisher
NII資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
dcmi資源タイプ	text
DCTERMS.bibliographicCitation	Journal of Biological Chemistry.268(14) P.10588-P.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	英語
DCTERMS.issued" scheme="DCTERMS.W3CDTF	1993-05
DC.identifier" scheme="DCTERMS.URI	http://hdl.handle.net/11094/71440
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.
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	英語
citation_date	1993-05
citation_journal_title	Journal of Biological Chemistry
citation_volume	268
citation_issue	14
citation_firstpage	10588
citation_lastpage	10592
citation_issn	1083351X
citation_public_url	http://hdl.handle.net/11094/71440

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