title: Detection of a local interaction of hen lysozyme under highly denaturing conditions using chemically 13C-enriched methionine resonance.
abstract: Using hen lysozyme in which the epsilon-carbons of two methionine residues are enriched with 13C nuclei, we found that there is a subtle difference in the chemical shift of the epsilon-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 degrees 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 epsilon-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.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka.
authors.forename: Y
authors.initials: Y
authors.lastname: Abe
authors.forename: T
authors.initials: T
authors.lastname: Ueda
authors.forename: K
authors.initials: K
authors.lastname: Kawano
authors.forename: Y
authors.initials: Y
authors.lastname: Tanaka
authors.forename: T
authors.initials: T
authors.lastname: Imoto
mesh_terms: D000478:Alkylation; D000595:Amino Acid Sequence; D000818:Animals; D002247:Carbon Isotopes; D002645:Chickens; D008715:Methionine / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry* / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D017510:Protein Folding; D013696:Temperature
chemical_list: D002247:Carbon Isotopes; D008715:Methionine; D009113:Muramidase