title: Iodotyrosine deiodinase is the first mammalian member of the NADH oxidase/flavin reductase superfamily.
abstract: The enzyme responsible for iodide salvage in the thyroid, iodotyrosine deiodinase, was solubilized from porcine thyroid microsomes by limited proteolysis with trypsin. The resulting protein retained deiodinase activity and was purified using anion exchange, dye, and hydrophobic chromatography successively. Peptide sequencing of the final isolate identified the gene responsible for the deiodinase. The amino acid sequence of the porcine enzyme is highly homologous to corresponding genes in a variety of mammals including humans, and the mouse gene was expressed in human embryonic kidney 293 cells to confirm its identity. The amino acid sequence of the deiodinase suggests the presence of three domains. The N-terminal domain provides a membrane anchor. The intermediate domain contains the highest sequence variability and lacks homology to structural motifs available in the common databases. The C-terminal domain is highly conserved and resembles bacterial enzymes of the NADH oxidase/flavin reductase superfamily. A three-dimensional model of the deiodinase based on the coordinates of the minor nitroreductase of Escherichia coli indicates that a Cys common to all of the mammal sequences is located adjacent to bound FMN. However, the deiodinase is not structurally related to other known flavoproteins containing redox-active cysteines or the iodothyronine deiodinases containing an active site selenocysteine.
authors.affiliation: Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20742, USA.
authors.forename: Jessica E
authors.initials: JE
authors.lastname: Friedman
authors.forename: James A
authors.initials: JA
authors.lastname: Watson
authors.forename: David W-H
authors.initials: DW
authors.lastname: Lam
authors.forename: Steven E
authors.initials: SE
authors.lastname: Rokita
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D003545:Cysteine; D038181:FMN Reductase / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D006801:Humans; D007453:Iodide Peroxidase / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D008861:Microsomes / Q000201:enzymology; D017434:Protein Structure, Tertiary; D017421:Sequence Analysis; D013552:Swine; D013961:Thyroid Gland / Q000201:enzymology
chemical_list: D007453:Iodide Peroxidase; D038181:FMN Reductase; D003545:Cysteine