The respiratory system of Pseudomonas putida: participation of cytochromes in electron transport.
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Biomedical subjects
Publications and source records attributed to W J Sweet.
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Nitrogenase activities and the patterns of in vivo inhibition of nitrogenase by NH+4 were compared in Rhodospirillum rubrum grown under several conditions of nitrogen availability. In cells grown on N2 or glutamate plus N2, nitrogenase activity was relatively low and was totally inhibited by added NH+4 in 15 to 20 min. In contrast, cells grown on glutamate alone displayed higher nitrogenase activity, and NH+4 had very little effect. Cells grown on limiting amounts of NH+4 had lower nitrogenase activity, but NH+4 produced little inhibitory effect. Uptake of NH+4 could be demonstrated under all of these conditions, and this uptake was blocked by DL-methionine-dl-sulfoximine. The data indicated that cells not recently exposed to NH+4 had no mechanism for rapidly turning off nitrogenase activity in response to sudden additions of NH+4. In contrast, cells grown in the presence of N2, which form NH+4 internally, inhibited nitrogenase activity relatively quickly in response to added NH+4.
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Optical absorbance difference spectra of membrane vesicles prepared from aerobically grown Pseudomonas putida indicated that, when harvested in logarithmic phase, the cells contained one c-type cytochrome and two or three b-type cytochromes, one of which was cytochrome o. As the cells grew into stationary phase and the oxygen concentration of the medium dropped to essentially zero, an additional component believed to be cytochrome d was produced. Both the o- and d-type cytochromes might function as terminal oxidases. No a-type cytochromes could be detected at any stage of growth. Polarographic measurement of oxygen utilization revealed that cyanide and azide are effective inhibitors of the oxidation of ascorbate coupled with 2,6-dichlorophenolindophenol or N,N,N',N'-tetramethyl-p-phenylenediamine in respiratory particles from either log-phase or stationary-phase cells. Reduced nicotinamide adenine dinucleotide- or succinate-dependent oxygen utilization, however, was sensitive to these inhibitors only in log-phase particles. These results indicate that an alternate terminal oxidase may be synthesized by this organism in response to restricted oxygen availability and that branching of the respiratory system may result.