title: Elucidation of the relationship between enzyme activity and internal motion using a lysozyme stabilized by cavity-filling mutations.
abstract: We investigated the activity and the internal motions of a stabilized mutant hen lysozyme (HEL) in which the residues M12 and L56 were mutated to L and F, respectively (LF mutant HEL). The result of the activity measurements against glycol chitin at various temperatures suggested that the temperature dependence of the activity of LF mutant HEL shifted to the high-temperature side compared with that of wild-type HEL. The detailed internal motions of LF mutant HEL in the absence and presence of a substrate analogue, (NAG)3, were examined by model-free analysis at 35 degrees C. The results showed that the internal motions of LF mutant HEL in the presence of (NAG)3 were drastically restricted compared with those in wild-type HEL. Our findings thus suggested that the mutation to the stabilized lysozyme restricted internal motions required for the enzymatic reaction.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Yoshida
authors.forename: T
authors.initials: T
authors.lastname: Ohkuri
authors.forename: S
authors.initials: S
authors.lastname: Kino
authors.forename: T
authors.initials: T
authors.lastname: Ueda
authors.forename: T
authors.initials: T
authors.lastname: Imoto
mesh_terms: D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D002645:Chickens; D002686:Chitin / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D004795:Enzyme Stability / Q000235:genetics; D005838:Genotype; D007700:Kinetics; D008958:Models, Molecular; D009113:Muramidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D019906:Nuclear Magnetic Resonance, Biomolecular; D017434:Protein Structure, Tertiary; D013696:Temperature
chemical_list: D002686:Chitin; C033828:N-acetylglucosaminitol; C031610:glycolchitin; D009113:Muramidase; D000117:Acetylglucosamine