title: The catalytic mechanism of decarboxylative hydroxylation of salicylate hydroxylase revealed by crystal structure analysis at 2.5 Å resolution.
abstract: The X-ray crystal structure of a salicylate hydroxylase from Pseudomonas putida S-1 complexed with coenzyme FAD has been determined to a resolution of 2.5 Å. Structural conservation with p- or m-hydroxybenzoate hydroxylase is very good throughout the topology, despite a low amino sequence identity of 20-40% between these three hydroxylases. Salicylate hydroxylase is composed of three distinct domains and includes FAD between domains I and II, which is accessible to solvent. In this study, which analyzes the tertiary structure of the enzyme, the unique reaction of salicylate, i.e. decarboxylative hydroxylation, and the structural roles of amino acids surrounding the substrate, are considered.
authors.affiliation: Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan; Research Reactor Institute, Kyoto University, Kumatori, Osaka 590-0494, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Uemura
authors.affiliation: Research Reactor Institute, Kyoto University, Kumatori, Osaka 590-0494, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Kita
authors.affiliation: Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan; Research Reactor Institute, Kyoto University, Kumatori, Osaka 590-0494, Japan.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Watanabe
authors.affiliation: Quantum Beam Science Center, Japan Atomic Energy Agency, Ibaraki 319-1195, Japan.
authors.forename: Motoyasu
authors.initials: M
authors.lastname: Adachi
authors.affiliation: Quantum Beam Science Center, Japan Atomic Energy Agency, Ibaraki 319-1195, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Kuroki
authors.affiliation: Research Reactor Institute, Kyoto University, Kumatori, Osaka 590-0494, Japan. Electronic address: morimoto@rri.kyoto-u.ac.jp.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Morimoto
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002384:Catalysis; D003198:Computer Simulation; D004789:Enzyme Activation; D006868:Hydrolysis; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000648:ultrastructure*; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: D006899:Mixed Function Oxygenases; C022619:salicylate 1-monooxygenase
keywords: Decarboxylative hydroxylation; Enzyme; Flavin-dependent; Salicylate hydroxylase; X-ray analysis