title: The copper site in nitrous oxide reductase.
abstract: The properties of the novel copper enzyme nitrous oxide reductase from denitrifying Pseudomonas stutzeri are described. Multifrequency electron paramagnetic resonance spectroscopy is used to characterize the various forms of the enzyme. The features observed at 2.4, 3.4, 4.5, 9.31 and 35 GHz are explained by a mixed-valence [Cu(1.5)...Cu(1.5)] S = 1/2 species with the unpaired electron delocalized between the two Cu nuclei. This site is also present in the catalytically inactive derivative of nitrous oxide reductase which was obtained from a transposon Tn5-induced mutant with defective chromophore biosynthesis. The resemblance of the low-frequency electron paramagnetic resonance spectra to the spectra for the so-called CuA of cytochrome c oxidase can be taken as a first indication that the CuA may have a structural and electronic arrangement similar to the electron-paramagnetic-resonance-detectable copper in nitrous oxide reductase. Results from oxidation/reduction experiments, and from a quantitative determination of sulfhydryl and disulfide residues in the various forms of nitrous oxide reductase, suggest the involvement of the redox-couple cysteine/cystine in the structural organization of the active site of nitrous oxide reductase.
authors.affiliation: Fakultät für Biologie, Universität Konstanz, Federal Republic of Germany.
authors.forename: P M
authors.initials: PM
authors.lastname: Kroneck
authors.forename: J
authors.initials: J
authors.lastname: Riester
authors.forename: W G
authors.initials: WG
authors.lastname: Zumft
authors.forename: W E
authors.initials: WE
authors.lastname: Antholine
mesh_terms: D001665:Binding Sites; D003300:Copper / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D003576:Electron Transport Complex IV / Q000378:metabolism; D008956:Models, Chemical; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D013438:Sulfhydryl Compounds; D003300:Copper; D010088:Oxidoreductases; C022763:nitrous oxide reductase; D003576:Electron Transport Complex IV