1H-NMR study of the intramolecular interaction of a substrate analogue covalently attached to aspartic acid-101 in lysozyme

Tadashi Ueda, Yoshimasa Isakari, Hidenori Aoki, Takanori Yasukochi, Shun ichi Masutomo, Keiichi Kawano, Yoshihiro Terada, Hidenori Yamada, Taiji Imoto

研究成果: ジャーナルへの寄稿記事

3 引用 (Scopus)

抄録

We prepared the lysozyme derivative in which the β-carboxyl group of Asp101 was modified with α-O-methyl N-glycylglucosaminide as an amide by means of the carbodi-imide reaction (α-MGG lysozyme). Since Asp101 residue is located at the edge of the active site cleft, a 1H-NMR study was carried out for this derivative in order to investigate the interaction between the introduced substituent and the active site cleft. It was confirmed that the α-MGG moiety sat in the active site cleft in α-MGG lysozyme from the reduction of line broadening of the NH-proton of Trp63 located in the active site cleft, the remarkable chemical shift change of the methyl group of the α-MGG moiety upon adding a trimer of N-acetyl-D-glucosamine [(NAG)3], and the NOE between the C6-proton resonance of Trp63 and the methyl resonance of the α-MGG moiety. Furthermore, α-MGG lysozyme had increased thermal stability compared with native lysozyme. Therefore, it was concluded that the α-MGG moiety covalently attached to Asp101 interacted with the active site cleft to increase the thermal stability of lysozyme.

元の言語英語
ページ(範囲)690-698
ページ数9
ジャーナルJournal of Biochemistry
109
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
Aspartic Acid
Nuclear magnetic resonance
Catalytic Domain
Substrates
Protons
Thermodynamic stability
Hot Temperature
Imides
Derivatives
Acetylglucosamine
Chemical shift
Amides
Proton Magnetic Resonance Spectroscopy

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

1H-NMR study of the intramolecular interaction of a substrate analogue covalently attached to aspartic acid-101 in lysozyme. / Ueda, Tadashi; Isakari, Yoshimasa; Aoki, Hidenori; Yasukochi, Takanori; Masutomo, Shun ichi; Kawano, Keiichi; Terada, Yoshihiro; Yamada, Hidenori; Imoto, Taiji.

:: Journal of Biochemistry, 巻 109, 番号 5, 01.01.1991, p. 690-698.

研究成果: ジャーナルへの寄稿記事

Ueda, T, Isakari, Y, Aoki, H, Yasukochi, T, Masutomo, SI, Kawano, K, Terada, Y, Yamada, H & Imoto, T 1991, '1H-NMR study of the intramolecular interaction of a substrate analogue covalently attached to aspartic acid-101 in lysozyme', Journal of Biochemistry, 巻. 109, 番号 5, pp. 690-698. https://doi.org/10.1093/oxfordjournals.jbchem.a123442
Ueda, Tadashi ; Isakari, Yoshimasa ; Aoki, Hidenori ; Yasukochi, Takanori ; Masutomo, Shun ichi ; Kawano, Keiichi ; Terada, Yoshihiro ; Yamada, Hidenori ; Imoto, Taiji. / 1H-NMR study of the intramolecular interaction of a substrate analogue covalently attached to aspartic acid-101 in lysozyme. :: Journal of Biochemistry. 1991 ; 巻 109, 番号 5. pp. 690-698.
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abstract = "We prepared the lysozyme derivative in which the β-carboxyl group of Asp101 was modified with α-O-methyl N-glycylglucosaminide as an amide by means of the carbodi-imide reaction (α-MGG lysozyme). Since Asp101 residue is located at the edge of the active site cleft, a 1H-NMR study was carried out for this derivative in order to investigate the interaction between the introduced substituent and the active site cleft. It was confirmed that the α-MGG moiety sat in the active site cleft in α-MGG lysozyme from the reduction of line broadening of the NH-proton of Trp63 located in the active site cleft, the remarkable chemical shift change of the methyl group of the α-MGG moiety upon adding a trimer of N-acetyl-D-glucosamine [(NAG)3], and the NOE between the C6-proton resonance of Trp63 and the methyl resonance of the α-MGG moiety. Furthermore, α-MGG lysozyme had increased thermal stability compared with native lysozyme. Therefore, it was concluded that the α-MGG moiety covalently attached to Asp101 interacted with the active site cleft to increase the thermal stability of lysozyme.",
author = "Tadashi Ueda and Yoshimasa Isakari and Hidenori Aoki and Takanori Yasukochi and Masutomo, {Shun ichi} and Keiichi Kawano and Yoshihiro Terada and Hidenori Yamada and Taiji Imoto",
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T1 - 1H-NMR study of the intramolecular interaction of a substrate analogue covalently attached to aspartic acid-101 in lysozyme

AU - Ueda, Tadashi

AU - Isakari, Yoshimasa

AU - Aoki, Hidenori

AU - Yasukochi, Takanori

AU - Masutomo, Shun ichi

AU - Kawano, Keiichi

AU - Terada, Yoshihiro

AU - Yamada, Hidenori

AU - Imoto, Taiji

PY - 1991/1/1

Y1 - 1991/1/1

N2 - We prepared the lysozyme derivative in which the β-carboxyl group of Asp101 was modified with α-O-methyl N-glycylglucosaminide as an amide by means of the carbodi-imide reaction (α-MGG lysozyme). Since Asp101 residue is located at the edge of the active site cleft, a 1H-NMR study was carried out for this derivative in order to investigate the interaction between the introduced substituent and the active site cleft. It was confirmed that the α-MGG moiety sat in the active site cleft in α-MGG lysozyme from the reduction of line broadening of the NH-proton of Trp63 located in the active site cleft, the remarkable chemical shift change of the methyl group of the α-MGG moiety upon adding a trimer of N-acetyl-D-glucosamine [(NAG)3], and the NOE between the C6-proton resonance of Trp63 and the methyl resonance of the α-MGG moiety. Furthermore, α-MGG lysozyme had increased thermal stability compared with native lysozyme. Therefore, it was concluded that the α-MGG moiety covalently attached to Asp101 interacted with the active site cleft to increase the thermal stability of lysozyme.

AB - We prepared the lysozyme derivative in which the β-carboxyl group of Asp101 was modified with α-O-methyl N-glycylglucosaminide as an amide by means of the carbodi-imide reaction (α-MGG lysozyme). Since Asp101 residue is located at the edge of the active site cleft, a 1H-NMR study was carried out for this derivative in order to investigate the interaction between the introduced substituent and the active site cleft. It was confirmed that the α-MGG moiety sat in the active site cleft in α-MGG lysozyme from the reduction of line broadening of the NH-proton of Trp63 located in the active site cleft, the remarkable chemical shift change of the methyl group of the α-MGG moiety upon adding a trimer of N-acetyl-D-glucosamine [(NAG)3], and the NOE between the C6-proton resonance of Trp63 and the methyl resonance of the α-MGG moiety. Furthermore, α-MGG lysozyme had increased thermal stability compared with native lysozyme. Therefore, it was concluded that the α-MGG moiety covalently attached to Asp101 interacted with the active site cleft to increase the thermal stability of lysozyme.

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