title: Extended heme promiscuity in the cyanobacterial cytochrome c oxidase: characterization of native complexes containing hemes A, O, and D, respectively.
abstract: The cyanobacteria Anacystis nidulans (Synechococcus sp. PCC6301), Synechocystis sp. PCC6803, Anabaena sp. PCC 7120, and Nostoc sp. PCC8009 were grown photoautotrophically under reduced oxygen tension in a medium with sulfate replaced by thiosulfate and nitrate replaced by ammonium as the S- and N-sources, respectively. In addition, Anabaena and Nostoc were grown under dinitrogen-fixing conditions in a medium free of combined nitrogen. Membranes were isolated from late-logarithmic cells (culture density corresponding to approximately 3 microliters packed cells per milliliter); cytoplasmic and thylakoid membranes were separated and purified according to established procedures. Acid-labile hemes were extracted from the membranes and subjected to reversed-phase high-performance liquid chromatography. Separated hemes were analyzed spectroscopically and identified by comparison with authentic standards. In addition to hemes B, A, and O, the latter of which was induced under semianaerobic conditions only, substitution of thiosulfate and ammonium for the oxy-anions sulfate and nitrate led to the appearance of spectrally discernible heme D in the membranes and extracts therefrom. However, spectroscopic and kinetic investigation of the membrane-bound heme D rather disproved any reaction with oxygen or carbon monoxide. Kinetic measurements performed with the membrane-bound respiratory oxidase gave evidence for only two kinetically competent terminal oxidases, a3 and o3, both apparently associated with a single type of apoprotein, viz. subunit I of the known cyanobacterial aa3-type cytochrome c oxidase. The heme D, on the other hand, seems to form a spectrally distinguished, yet kinetically ill-defined hemoprotein complex which does not qualify as a fully functional d-type terminal oxidase on our (wild-type) cyanobacteria even after growth under semianaerobic pseudo-reducing conditions. Also growth (of Anabaena and Nostoc) under dinitrogen-fixing conditions did not change this situation. Thus, we are left with (wild-type) cyanobacteria forming an unbranched respiratory chain with only a single type of terminal oxidase protein, viz. the known aa3-type cytochrome c oxidase. This oxidase, however, may incorporate different prosthetic (heme) groups in the sense of "heme promiscuity." Biosynthesis of the different heme groups thereby seems to respond to the ambient redox environment. In particular, however, conditions for expression of the two quinol oxidases potentially and additionally coded for by the genome of, e. g., Synechocystis sp. PCC6803 (see http://www.kazusa.or.jp/cyano), have not yet been found.
authors.affiliation: Molecular Bioenergetics Group, Institute of Physical Chemistry, University of Vienna, Wien, A-1090, Austria.
authors.forename: S
authors.initials: S
authors.lastname: Fromwald
authors.forename: R
authors.initials: R
authors.lastname: Zoder
authors.forename: M
authors.initials: M
authors.lastname: Wastyn
authors.forename: M
authors.initials: M
authors.lastname: Lübben
authors.forename: G A
authors.initials: GA
authors.lastname: Peschek
mesh_terms: D000143:Acids / Q000378:metabolism; D015153:Blotting, Western; D002248:Carbon Monoxide / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D000458:Cyanobacteria / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D004227:Dithionite / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000737:chemistry*; D006418:Heme / Q000031:analogs & derivatives / Q000032:analysis* / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007425:Intracellular Membranes / Q000201:enzymology; D009243:NAD / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D013057:Spectrum Analysis; D013885:Thiosulfates / Q000378:metabolism
chemical_list: D000143:Acids; D000644:Quaternary Ammonium Compounds; D013885:Thiosulfates; D009243:NAD; C069808:heme O; D004227:Dithionite; C023647:pyridine hemochrome; C027728:heme a; D006418:Heme; C080647:heme d; D002248:Carbon Monoxide; D006861:Hydrogen Peroxide; D003576:Electron Transport Complex IV; D010100:Oxygen