Preparation and properties of a lysozyme derivative in which two domains are cross-linked intramolecularly between trp62 and asp101

Tadashi Ueda, Hidenori Yamada, Noriko Sakamoto, Yoshito Abe, Keiichi Kawano, Yoshihiro Terada, Taiji Imoto

研究成果: ジャーナルへの寄稿記事

19 引用 (Scopus)

抄録

A lysozyme derivative in which two domains were cross-linked intramolecularly was newly prepared by means of a two-step reaction. First,the β-carboxyl group of Asp1O1 in lysozyme was selectively modified with 2-(2-pyridyldithio)ethylamine in the presence of l-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride. After reduction of the pyridyldithio moiety of Asp101 modified lysozyme at pH4.5 with dithiothreitol, the derivative was allowed to cross-link intramolecularly by reaction with 1,3-dichloroace-tone at pH 7. Intramolecularly cross-linked lysozyme thus formed was purified by gel chromatography followed by ion-exchange chromatography. Based on the results of1H-NMR and peptide analyses, it was concluded that Asp101 was cross-linked to Trp62 with a -CH2COCH2SCH2CH2NH- bridge in this derivative. The derivative showed minor but distinct activity against Micrococcus lysodeikticus and glycol chitin. Its melting temperature for thermal denaturation was higher by 7.3°C than that of native lysozyme at pH3.

元の言語英語
ページ(範囲)719-725
ページ数7
ジャーナルJournal of biochemistry
110
発行部数5
DOI
出版物ステータス出版済み - 1 1 1991

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Muramidase
Derivatives
Chromatography
Carbodiimides
Micrococcus
Denaturation
Dithiothreitol
Ion Exchange Chromatography
Freezing
Gel Chromatography
Melting point
Ion exchange
Hot Temperature
Gels
Nuclear magnetic resonance
Peptides
Temperature

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

Preparation and properties of a lysozyme derivative in which two domains are cross-linked intramolecularly between trp62 and asp101. / Ueda, Tadashi; Yamada, Hidenori; Sakamoto, Noriko; Abe, Yoshito; Kawano, Keiichi; Terada, Yoshihiro; Imoto, Taiji.

:: Journal of biochemistry, 巻 110, 番号 5, 01.01.1991, p. 719-725.

研究成果: ジャーナルへの寄稿記事

Ueda, Tadashi ; Yamada, Hidenori ; Sakamoto, Noriko ; Abe, Yoshito ; Kawano, Keiichi ; Terada, Yoshihiro ; Imoto, Taiji. / Preparation and properties of a lysozyme derivative in which two domains are cross-linked intramolecularly between trp62 and asp101. :: Journal of biochemistry. 1991 ; 巻 110, 番号 5. pp. 719-725.
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abstract = "A lysozyme derivative in which two domains were cross-linked intramolecularly was newly prepared by means of a two-step reaction. First,the β-carboxyl group of Asp1O1 in lysozyme was selectively modified with 2-(2-pyridyldithio)ethylamine in the presence of l-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride. After reduction of the pyridyldithio moiety of Asp101 modified lysozyme at pH4.5 with dithiothreitol, the derivative was allowed to cross-link intramolecularly by reaction with 1,3-dichloroace-tone at pH 7. Intramolecularly cross-linked lysozyme thus formed was purified by gel chromatography followed by ion-exchange chromatography. Based on the results of1H-NMR and peptide analyses, it was concluded that Asp101 was cross-linked to Trp62 with a -CH2COCH2SCH2CH2NH- bridge in this derivative. The derivative showed minor but distinct activity against Micrococcus lysodeikticus and glycol chitin. Its melting temperature for thermal denaturation was higher by 7.3°C than that of native lysozyme at pH3.",
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AU - Abe, Yoshito

AU - Kawano, Keiichi

AU - Terada, Yoshihiro

AU - Imoto, Taiji

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N2 - A lysozyme derivative in which two domains were cross-linked intramolecularly was newly prepared by means of a two-step reaction. First,the β-carboxyl group of Asp1O1 in lysozyme was selectively modified with 2-(2-pyridyldithio)ethylamine in the presence of l-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride. After reduction of the pyridyldithio moiety of Asp101 modified lysozyme at pH4.5 with dithiothreitol, the derivative was allowed to cross-link intramolecularly by reaction with 1,3-dichloroace-tone at pH 7. Intramolecularly cross-linked lysozyme thus formed was purified by gel chromatography followed by ion-exchange chromatography. Based on the results of1H-NMR and peptide analyses, it was concluded that Asp101 was cross-linked to Trp62 with a -CH2COCH2SCH2CH2NH- bridge in this derivative. The derivative showed minor but distinct activity against Micrococcus lysodeikticus and glycol chitin. Its melting temperature for thermal denaturation was higher by 7.3°C than that of native lysozyme at pH3.

AB - A lysozyme derivative in which two domains were cross-linked intramolecularly was newly prepared by means of a two-step reaction. First,the β-carboxyl group of Asp1O1 in lysozyme was selectively modified with 2-(2-pyridyldithio)ethylamine in the presence of l-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride. After reduction of the pyridyldithio moiety of Asp101 modified lysozyme at pH4.5 with dithiothreitol, the derivative was allowed to cross-link intramolecularly by reaction with 1,3-dichloroace-tone at pH 7. Intramolecularly cross-linked lysozyme thus formed was purified by gel chromatography followed by ion-exchange chromatography. Based on the results of1H-NMR and peptide analyses, it was concluded that Asp101 was cross-linked to Trp62 with a -CH2COCH2SCH2CH2NH- bridge in this derivative. The derivative showed minor but distinct activity against Micrococcus lysodeikticus and glycol chitin. Its melting temperature for thermal denaturation was higher by 7.3°C than that of native lysozyme at pH3.

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