title: Rapid kinetics of dehalogenation promoted by iodotyrosine deiodinase from human thyroid.
abstract: Reductive dehalogenation such as that catalyzed by iodotyrosine deiodinase (IYD) is highly unusual in aerobic organisms but necessary for iodide salvage from iodotyrosine generated during thyroxine biosynthesis. Equally unusual is the dependence of this process on flavin. Rapid kinetics have now been used to define the basic processes involved in IYD catalysis. Time-dependent quenching of flavin fluorescence was used to monitor halotyrosine association to IYD. The substrates chloro-, bromo-, and iodotyrosine bound with similar rate constants (kon) ranging from 1.3 × 10(6) to 1.9 × 10(6) M(-1) s(-1). Only the inert substrate analogue fluorotyrosine exhibited a significantly (5-fold) slower kon (0.3 × 10(6) M(-1) s(-1)). All data fit a standard two-state model and indicated that no intermediate complex accumulated during closure of the active site lid induced by substrate. Subsequent halide elimination does not appear to limit reactions of bromo- and iodotyrosine since both fully oxidized the reduced enzyme with nearly equivalent second-order rate constants (7.3 × 10(3) and 8.6 × 10(3) M(-1) s(-1), respectively) despite the differing strength of their carbon-halogen bonds. In contrast to these substrates, chlorotyrosine reacted with the reduced enzyme approximately 20-fold more slowly and revealed a spectral intermediate that formed at approximately the same rate as the bromo- and iodotyrosine reactions.
authors.affiliation: †Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, United States.
authors.forename: Kostyantyn D
authors.initials: KD
authors.lastname: Bobyk
authors.affiliation: ‡Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States.
authors.forename: David P
authors.initials: DP
authors.lastname: Ballou
authors.affiliation: †Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, United States.|§Department of Chemistry, Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, United States.
authors.forename: Steven E
authors.initials: SE
authors.lastname: Rokita
mesh_terms: D055162:Biocatalysis; D020134:Catalytic Domain; D004138:Dinitrocresols / Q000737:chemistry; D006801:Humans; D007453:Iodide Peroxidase / Q000737:chemistry*; D007700:Kinetics; D007470:Monoiodotyrosine / Q000737:chemistry; D010084:Oxidation-Reduction; D011485:Protein Binding; D013961:Thyroid Gland / Q000201:enzymology*
chemical_list: D004138:Dinitrocresols; C024132:4,6-dinitro-o-cresol; D007453:Iodide Peroxidase; D007470:Monoiodotyrosine