Nucleotide exchange in membrane vesicles from the photosynthetic bacterium Rhodopseudomonas capsulata

Membrane vesicles (heavy chromatophores) prepared from the photosynthetic bacteria Rhodopseudomonas capsulata catalyze photophosphorylation of exogenous ADP and also take up [³H]ADP from the external medium. The rate of uptake depends on the concentration of external ADP reaching half-maximal velocity at 2.7 mm. The rate increases also with the increase in the concentration of internal ADP. Vesicles, preloaded with [³H]ADP release the radioactive nucleotide when ADP is included in the external medium. Regular chromatophores, which are inside-out membrane vesicles also take up [³H]ADP from the external medium when preloaded with ADP. These results are interpreted to indicate the existence of nucleotide transport across the cytoplasmic membrane of these bacteria which is catalyzed by an ADP exchange carrier.     

Respiration in air and water of four mediterranean trochids

Data on the morphology and behavior of five species of spionid polychaete from muddy sand habitats in both the United Kingdom and the northwest coast of North America are presented. The data are consistent with the hypothesis that browsing predation by visual predators is important now and has been important in the evolutionary past. As predicted by such a hypothesis, regenerating individuals are common in all the populations examined. Individuals belonging to species that expose their anterior ends while feeding and/or defecating have cryptic anteriors (Pygospio elegans Claparède, Rhynchospio glutaeus (Ehlers), Malacoceros fuliginosus (Claparède), and Spiophanes bombyx (Claparède)). Individuals belonging to species that do not expose their anteriors while feeding and/or defecating do not have cryptic anteriors (Pseudopolydora kempi (Southern)). Given these data it is still unclear as to how many of the geographic patterns of coloration that have been reported for infauna are associated with the presence of visual predation. Further investigations are suggested.     

Effect of oxygen on acetylene reduction by photosynthetic bacteria

The effect of dissolved oxygen concentration on nitrogenase activity was studied in three species of photosynthetic bacteria. The O2 concentration in the cell suspension was measured with an O2 electrode inserted into the reaction vessel. Acetylene reduction by whole cells of Rhodopseudomonas capsulata, Rhodospirillum rubrum, and Chromatium vinosum strain D was inhibited 50% by 0.73, 0.32, and 0.26 microM O2, respectively. The inhibition of the activity by O2 in R. capsulata usually was reversed completely by reestablishing anaerobic conditions. In R. rubrum and C. vinosum the inhibition was only partially reversible. The respiration rate of R. capsulata was the highest of the three, that of R. rubrum was intermediate, and that of C. vinosum was lowest. R. capsulata and R. rubrum cells were broken after their acetylene reduction activity in vivo had been completely inhibited by O2, and nitrogenase was found to be active in vitro. A concentration of cyanide that did not affect acetylene reduction activity, but which inhibited 75 to 90% of the O2 uptake by whole cells of R. capsulata, shifted the O2 concentration causing 50% inhibition of nitrogenase activity from 0.73 microM to 2.03 microM. These results are in accordance with the assumption that within a limited range of O2 concentrations, the respiratory activity of the cells is enough to scavenge the O2 and to keep the interior of the cells essentially anaerobic. It is suggested that O2 inhibits nitrogenase activity by competing for a limited supply of electrons. When cyanide is present, respiration is slower but is adequate to keep the nitrogenase environment in the cell anaerobic. The lower respiration rate may allow a greater proportion of the electrons to be used for acetylene reduction.     

Distribution and movement of toxaphene in anaerobic saline marsh soils

The biological oxygen demand (BOD) of filtered water from Lake Wingra, Wisconsin is significantly higher in the littoral zone than in the pelagial zone. Laboratory experiments indicate that BOD is not influenced by water temperature at the time of sampling or by enrichment with nitrate or ammonia. Rather, enrichment with macrophyte leachate sharply increases BOD, and enrichment with phosphate produces a small but significant increase in BOD. We conclude that high BOD in littoral waters of the lake is an indication of production of labile organic matter in the water by dense beds of the macrophyte Myriophyllum spicatum.     

Production and characterization of antisera against three individual NHC proteins; a case of a generally distributed NHC protein

In order to assess the selectivity of the distribution patterns of individual nonhistone chromosomal proteins (NHC proteins), immunofluorescent staining experiments were performed on Drosophila polytene chromosomes. Antisera have been prepared against three individual NHC proteins which were isolated by sequential preparative slab gel isoelectric focusing and SDS polyacrylamide gel electrophoresis. In two cases, immunofluorescent staining of the chromosomes indicated a specific limited distribution pattern; apparently the antigen in each case is present at a reproducible and distinct subset of chromomeres. This type of pattern has also been obtained with antisera prepared against molecular weight subfractions of NHC proteins (Silver and Elgin, 1977). Each selective fluorescence distribution pattern obtained so far is reproducible and unique to the antiserum under study. In a third case, an antiserum caused prominant staining at dense chromomeres and the chromocenter in a pattern mimicking DNA (and presumably histone) distribution. Indirect radioimmunostaining of SDS and isoelectric focusing gels on which total NHC proteins had been separated confirmed that this antiserum reacted specifically with a protein(s) of molecular weight 21,000 D and pI 5.2. The data in conjunction with absorption experiments indicates that the chromosomal staining is due to an interaction of antibodies with NHC protein(s) and not with histones. This finding suggests that at least one major acidic NHC protein plays a very general role (comparable to that of the histones) in maintaining chromatin structure.     

A requirement for cytoplasmic protein synthesis during chloroplast senescence in the aquatic plant Anacharis canadensis

Cycloheximide (CHI) at 1 µg/liter delayed the loss ofchlorophyll from detached Anacharis canadensis leaflets senescingin the dark. Chloramphenicol (CAP) and streptomycin (SM) slightlyaccelerated the loss. CHI was effective even during the laterstages of senescence in preventing further loss of chlorophyll.Senescence proceeded normally upon return of the leaflets intowater. The need for cytoplasmic protein synthesis during chloroplastsenescence and the types of proteins involved are discussed. (Received March 26, 1976; )     

The location and function of cytochrome c2 in Rhodopseudomonas capsulate membranes.

Two fractions of membrane preparations, a heavy and a light one were isolated from mildly broken Rhodopseudomonas capsulata cells. The light fraction which contained vesicles similar to the regular chromatophores obtained by sonication and a heavy fraction which appeared in electron micrographs to consist of cell fragments which were designated as heavy chromatophores and were composed of broken cell envelopes containing closely packed vesicles enclosed within the cytoplasmic membrane. Both types of chromatophores catalyzed photophosphorylation. However, cytochrome c2 could be washed out only from the heavy chromatophores. Photophosphorylation activity which was lost by the removal of the cytochrome could be restored by addition of either cytochrome c2 or phenazine methosulphate. Light induced proton efflux in heavy chromatophores in contrast to proton influx in regular chromatophores. The washed heavy chromatophores did not lose the light induced proton movement. Light induced quenching of 9-aminoacridine and atebrin fluorescence in chromatophores, while the fluorescence was enhanced in the heavy chromatophores. The washing did not affect the fluorescence changes of the heavy chromatophores but caused a reduction of the steady state of the carotenoid absorbance shift. It is suggested that the membrane in the heavy chromatophores is oriented inside out with respect to the membrane in regular chromatophores. Cytochrome c2 which is attached to that side of the membrane facing the outside medium could be removed from the heavy chromatophors and reconstituted to them. The role of cytochrome c2 in photophosphorylation is discussed.