Photosystem-I-driven inorganic carbon transport in the cyanobacterium, Anacystis nidulans

Ogawa, Teruo; Miyano, Akira; Inoue, Yorinao

doi:10.1016/0005-2728(85)90029-5

Cited by 103 publications

(62 citation statements)

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“…A high C02-requiring mutant (El) of Anacystis nidulans (7) and the wild type (treated with IAc, an inhibitor of CO2 fixation) were used in these experiments. Uptake of CO2 from the medium at air level of CO2 by this mutant (4,7) or by the wild type in the presence of IAc (1 1,12) represents the build up of the intracellular C, pool. The results demonstrated a one to one stoichiometry between the amounts of H+ appearing in the medium and CO2 taken up by the cells, even under the conditions where CO2 is transported.…”

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confidence: 99%

The Stoichiometry between CO₂ and H⁺ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

Ogawa¹,

Kaplan²

1987

Plant Physiol.

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The pH of the medium during CO2 uptake into the intracellular inorganic carbon (CQ) pool of a high C02-requiring mutant (Ej) and wild type of Anacystis nidulans R2 was measured. Experiments were performed under conditions where photosynthetic CO2 fixation is inhibited. There was an acidification of the medium during CO2 uptake in the light and an alkalization during CO2 efflux after darkening. A one to one stoichiometry existed between the amounts of H' appearing in the medium and CO2 taken up into the intracellular Ci pool, regardless of the carbon species transported. The results indicate that (a) CO2 is taken up simultaneously with an efflux of equimolar H+, probably produced as a result of CO2 hydration during transport and (b) HC03-produced by hydration of CO2 in the medium was transported into the cells without accompanying net flux of Ho or OH-. The influx and efflux of Ci during Ci transport produced nonequilibrium between CO2 and HCO3-in the medium, with the concentration of HC03-being higher than that expected under equilibrium conditions. The nonequilibrium was present even under the conditions where the influx of Ci is compensated by its efflux. The direction of this nonequilibrium suggested that efflux of HCO3-occurs during uptake of Ci.Cyanobacteria are capable of accumulating Ci2 internally and thus possess a 'C02 concentrating mechanism ' ( 1, 3, 6, 9, 10 (8,15) and HC03-appears to be the species which arrives at the inner membrane surface, regardless of the species supplied (15). Since Ci accumulated within the cells is mostly HCO3-, uptake of CO2 during the accumulation of Ci would be accompanied by an influx of OH-(or H+ efflux). Stoichiometry between H+ flux and CO2 taken up into the intracellular C, pool, however, has not been measured. When HC03-is supplied to the cell suspension in a closed system, the photosynthetic utilization of C, leads to alkalization of the medium and a decrease of the extemal Ci concentration (2,9 gas-analysis system where CO2 is supplied to (or taken up from) the cell suspension, it is possible to keep the steady-state C, concentration of the medium constant while measuring the CO2 exchange of the system. Furthermore, the use of mutants or inhibitors enables one to separate CO2 uptake into the intracellular Ci pool from CO2 fixation into acid-stable products.In this study, we measured the flux of H+ during CO2 uptake in the light and CO2 efflux after darkening, using an open gasanalysis system. A high C02-requiring mutant (El) of Anacystis nidulans (7) and the wild type (treated with IAc, an inhibitor of CO2 fixation) were used in these experiments. Uptake of CO2 from the medium at air level of CO2 by this mutant (4, 7) or by the wild type in the presence of IAc (1 1, 12) represents the build up of the intracellular C, pool. The results demonstrated a one to one stoichiometry between the amounts of H+ appearing in the medium and CO2 taken up by the cells, even under the conditions where CO2 is transported. The concentration ratio of the different Ci specie...

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confidence: 99%

The Stoichiometry between CO₂ and H⁺ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

Ogawa¹,

Kaplan²

1987

Plant Physiol.

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“…The C02-concentrating mechanism involves an active C, transport, which is driven by PSI reaction of photosynthesis (20)(21)(22). While the presently available evidence does not allow one to explain the transport with a specific mechanism, three models are most commonly discussed: (a) a primary electrogenic HCO3 pump (9), (b) H+/HCO3r symport or OH-/HCO3 antiport secondary to an H+ extrusion pump (4,16), and (c) Na+/HCO3 symport secondary to a Na+/H+ exchange (8,17 (3,21).…”

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confidence: 99%

Biosynthesis of a 42-kD Polypeptide in the Cytoplasmic Membrane of the Cyanobacterium Anacystis nidulans Strain R2 during Adaptation to Low CO₂ Concentration

1986

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When cells of Anacystis nidulans strain R2 grown under high CO2 conditions (3%) were transferred to low CO2 conditions (0.05%), their ability to accumulate inorganic carbon (C1) increased up to 8 times.Cytoplasmic membranes (plasmalemma) isolated at various stages of low CO2 adaptation were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. There was a marked increase of a 42-kilodalton polypeptide in the cytoplasmic membrane during adaptation; a linear relationship existed between the amount of this polypeptide and the Cr accumulating capability of the cells. No significant changes were observed during this process in the amount of other polypeptides in the cytoplasmic membranes or in the polypeptide profiles of the thylakoid membranes, cell walls, and soluble fractions. Spectinomycin, an inhibitor of protein biosynthesis, inhibited both the increase of the 42-kilodalton polypeptide and the induction of high Ci-accumulating capability. The incorporation of 135SIsulfate into membrane proteins was greatly reduced during low CO2 adaptation. Radioautograms of the 35S-labeled membrane proteins revealed that synthesis of the 42-kilodalton polypeptide in the cytoplasmic membrane was specifically activated during the adaptation, while that of most other proteins was greatly suppressed. These results suggested that the 42-kilodalton polypeptide in the cytoplasmic membrane is involved in the active Ci transport by A. nidulans strain R2 and its synthesis under low CO2 conditions leads to high Cr-transporting activity. mechanism of Ci transport.Exposure of high C02-grown cells ofcyanobacteria to low CO2 conditions increases their Ci-accumulating capability and their photosynthetic affinity to C1 (15,25,28 Cyanobacteria have a mechanism to transport exogenous Cj2 and concentrate it internally (1,7,13,16,20,27). The C02-concentrating mechanism involves an active C, transport, which is driven by PSI reaction of photosynthesis (20)(21)(22). While the presently available evidence does not allow one to explain the transport with a specific mechanism, three models are most commonly discussed: (a) a primary electrogenic HCO3 pump (9), (b) (29) under aeration with 3% CO2 in air. Continuous illumination was provided by fluorescent lamps at 120 ,umol PAR/M2 -s. For low CO2 adaptation experiments, cells were harvested by centrifugation at 2200g for 10 min, resuspended in fresh BG-11 growth medium supplemented with 20 mM Hepes-NaOH buffer (pH 7.0), and aerated with air containing about 0.05% CO2 under the same light conditions. Gas Exchange Measurements. Cells were harvested by centrifugation at 3000g for 5 min, resuspended in 30 ml of 40 mM Hepes-NaOH buffer (pH 7.0 OMATA AND OGAWA 2* s); the postillumination CO2 burst was measured in the presence of 3 mm iodoacetamide which inhibits photosynthetic CO2 fixation completely (3,21). It has been shown that the amount of intracellular Ci calculated by the CO2 burst method agrees with that measured using the silicone oil-centrifugation method (23). The intrace...

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“…Cyanobacteria possess a C02-concentrating mechanism that involves active transport of C,2 driven by photosystem-I energy (2,9,10,13,16,19 …”

Section: Isolation Of Mutantsmentioning

confidence: 99%

“…Cyanobacteria possess a C02-concentrating mechanism that involves active transport of C,2 driven by photosystem-I energy (2,9,10,13,16,19). Both CO2 and HCO3-are transported into the cells by this mechanism, and HCO3-appears to be the species that arrives at the inner membrane surface regardless of the species supplied (25).…”

Section: Isolation Of Mutantsmentioning

confidence: 99%

Mutants of Synechocystis PCC6803 Defective in Inorganic Carbon Transport

Ogawa

1990

Plant Physiol.

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Eighty mutants of Synechocystis PCC6803 that require high CO2 for growth were examined with a mass spectrometer for their ability to take up CO2 in the light. Two of these mutants (type A) did not show any CO2 uptake while the rest of the mutants (type B) took up CO2 actively. Type A mutants (RKa and RKb) and one type B mutant (RK11) were partially characterized. At 3% C02, growth rates of the mutants and the wild type (WT) were similar.Under air levels of C02, growth of RKa and RKb was very slow, and RK11 did not grow at all. The photosynthetic affinities for inorganic carbon (Cl) in these three mutants were about 100 times lower than the affinity in WT. The following characteristics of type A mutants indicated that the mutants have a defect in their C02-transport system: (a) the activity of 13C1602 uptake in RKa and RKb in the light was less than 5% the activity in WT, and (b) each mutant had only a low level of activity of 14C02 uptake as measured by the method of silicone oil-filtering centrifugation. The HC03-transport system was also impaired in these mutants. The activity of H4C03-uptake MATERIALS AND METHODS Isolation of MutantsCyanobacteria possess a C02-concentrating mechanism that involves active transport of C,2 driven by photosystem-I energy (2,9,10,13,16,19

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Photosystem-I-driven inorganic carbon transport in the cyanobacterium, Anacystis nidulans

Cited by 103 publications

References 22 publications

The Stoichiometry between CO₂ and H⁺ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

The Stoichiometry between CO₂ and H⁺ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

Biosynthesis of a 42-kD Polypeptide in the Cytoplasmic Membrane of the Cyanobacterium Anacystis nidulans Strain R2 during Adaptation to Low CO₂ Concentration

Mutants of Synechocystis PCC6803 Defective in Inorganic Carbon Transport

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Photosystem-I-driven inorganic carbon transport in the cyanobacterium, Anacystis nidulans

Cited by 103 publications

References 22 publications

The Stoichiometry between CO2 and H+ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

The Stoichiometry between CO2 and H+ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

Biosynthesis of a 42-kD Polypeptide in the Cytoplasmic Membrane of the Cyanobacterium Anacystis nidulans Strain R2 during Adaptation to Low CO2 Concentration

Mutants of Synechocystis PCC6803 Defective in Inorganic Carbon Transport

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The Stoichiometry between CO₂ and H⁺ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

The Stoichiometry between CO₂ and H⁺ Fluxes Involved in the Transport of Inorganic Carbon in Cyanobacteria

Biosynthesis of a 42-kD Polypeptide in the Cytoplasmic Membrane of the Cyanobacterium Anacystis nidulans Strain R2 during Adaptation to Low CO₂ Concentration