Detection of a local interaction of hen lysozyme under highly denaturing conditions using chemically 13C-enriched methionine resonance

Yoshito Abe, Tadashi Ueda, Keiichi Kawano, Yoshitsugu Tanaka, Taiji Imoto

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3 Citations (Scopus)

Abstract

Using hen lysozyme in which the ε-carbons of two methionine residues are enriched with 13C nuclei, we found that there is a subtle difference in the chemical shift of the ε-carbon resonances between Met 12 and Met 105 in thermally denatured lysozyme without any reduction of disulfide bonds at pD 3.8, and also in reduced S-alkylated lysozyme at pD 3.8 and 35°C. The difference in the chemical shift was abolished on digestion with TPCK-trypsin and the chemical shifts of both resonances converged to that of Met 12, whose chemical shift is identical to that in the randomly coiled state. Therefore, it is suggested that the chemical shift in the ε-carbon resonance of Met 105 is different from that in the randomly coiled state due to an interaction involving Met 105. In order to locate the interaction involving Met 105, fragmentation of the reduced S-alkylated lysozyme into the peptides was carried out by means of chemical cleavage or specific endoprotease digestion. As a result, the local interaction of Met 105 or the residues around Met 105 with eleven residues at the C-terminus of lysozyme is suggested to occur.

Original languageEnglish
Pages (from-to)313-317
Number of pages5
JournalJournal of Biochemistry
Volume123
Issue number2
DOIs
Publication statusPublished - Jan 1 1998

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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