title: Observation of the pre-steady state process in thermolysin catalysis with a fluorescent displacement probe at low pH.
abstract: The pre-steady state process in the thermolysin-catalyzed hydrolysis of Cbz-Gly-Phe-Ala was observed at pH 4.5 by fluorescence stopped-flow method using Dns-Phe as a displacement probe. After the confirmation of the pre-equilibrium hypothesis for the binary interaction, the nonlinear substrate concentration dependence of the apparent kinetic constant for the pre-steady state process was analyzed and an existence of multi-intermediates was proposed.
authors.affiliation: Department of Polymer Science and Engineering, Kyoto Institute of Technology, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Kunugi
authors.forename: M
authors.initials: M
authors.lastname: Yokoyama
authors.forename: A
authors.initials: A
authors.lastname: Sakamoto
mesh_terms: D000595:Amino Acid Sequence; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D013820:Thermolysin / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D009842:Oligopeptides; D013820:Thermolysin