An improved method for preparing lysozyme with chemically 13C-Enriched methionine residues using 2-aminothiophenol as a reagent of thiolysis

Yoshito Abe, Tadashi Ueda, Taiji Imoto

研究成果: ジャーナルへの寄稿学術誌査読

2 被引用数 (Scopus)

抄録

Jones et al. have reported that the ε-carbons of methionine residues in myoglobin can be enriched with stable isotope (13C) in two steps, i.e., methylation of methionine residues with 13CH3I in the protein and thiolysis using dithiothreitol. Using their method, we failed to prepare active lysozyme in which the ε-carbons of methionine residues are enriched with 13C, because many side reactions took place under the thiolysis condition (pH 10.5, 37°C). When we employed 2-aminothiophenol as a reagent for thiolysis, the reduction proceeded under a weakly acidic condition to afford fully active lysozyme, in which the ε-carbons of two methionine residues were enriched with 13C, in a 30% yield. Analysis of the 13C-edited NOESY spectra of 13C-enriched methionine lysozyme in the absence and presence of a substrate analogue indicated the occurrence of conformational change around Met 105 in lysozyme.

本文言語英語
ページ(範囲)1153-1159
ページ数7
ジャーナルJournal of biochemistry
122
6
DOI
出版ステータス出版済み - 12月 1997
外部発表はい

!!!All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学

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