title: Identification and biochemical characterization of Arabidopsis thaliana sulfite oxidase. A new player in plant sulfur metabolism.
abstract: In mammals and birds, sulfite oxidase (SO) is a homodimeric molybdenum enzyme consisting of an N-terminal heme domain and a C-terminal molybdenum domain (EC ). In plants, the existence of SO has not yet been demonstrated, while sulfite reductase as part of sulfur assimilation is well characterized. Here we report the cloning of a plant sulfite oxidase gene from Arabidopsis thaliana and the biochemical characterization of the encoded protein (At-SO). At-SO is a molybdenum enzyme with molybdopterin as an organic component of the molybdenum cofactor. In contrast to homologous animal enzymes, At-SO lacks the heme domain, which is evident both from the amino acid sequence and from its enzymological and spectral properties. Thus, among eukaryotes, At-SO is the only molybdenum enzyme yet described possessing no redox-active centers other than the molybdenum. UV-visible and EPR spectra as well as apparent K(m) values are presented and compared with the hepatic enzyme. Subcellular analysis of crude cell extracts showed that SO was mostly found in the peroxisomal fraction. In molybdenum cofactor mutants, the activity of SO was strongly reduced. Using antibodies directed against At-SO, we show that a cross-reacting protein of similar size occurs in a wide range of plant species, including both herbacious and woody plants.
authors.affiliation: Botanical Institute, Technical University of Braunschweig, 38023 Braunschweig, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Eilers
authors.forename: G
authors.initials: G
authors.lastname: Schwarz
authors.forename: H
authors.initials: H
authors.lastname: Brinkmann
authors.forename: C
authors.initials: C
authors.lastname: Witt
authors.forename: T
authors.initials: T
authors.lastname: Richter
authors.forename: J
authors.initials: J
authors.lastname: Nieder
authors.forename: B
authors.initials: B
authors.lastname: Koch
authors.forename: R
authors.initials: R
authors.lastname: Hille
authors.forename: R
authors.initials: R
authors.lastname: Hänsch
authors.forename: R R
authors.initials: RR
authors.lastname: Mendel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000201:enzymology*; D015153:Blotting, Western; D002645:Chickens; D003067:Coenzymes; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D015723:Gene Library; D006418:Heme / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008667:Metalloproteins / Q000737:chemistry; D008969:Molecular Sequence Data; D000090006:Molybdenum Cofactors; D016366:Open Reading Frames; D010084:Oxidation-Reduction; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D020675:Peroxisomes / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011621:Pteridines / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism*; D013997:Time Factors; D014026:Nicotiana / Q000201:enzymology; D014466:Ultraviolet Rays
chemical_list: D003067:Coenzymes; D006418:Heme; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D050862:Oxidoreductases Acting on Sulfur Group Donors; D011621:Pteridines; D011994:Recombinant Proteins; D013455:Sulfur; C022261:molybdopterin