Active HCO3− transport in cyanobacteria

1991 ◽  
Vol 69 (5) ◽  
pp. 936-944 ◽  
Author(s):  
George S. Espie ◽  
Anthony G. Miller ◽  
Ramani A. Kandasamy ◽  
David T. Canvin
Keyword(s):  
Inorganic Carbon ◽  
Dissolved Inorganic Carbon ◽  
Reaction Medium ◽  
Transport Systems ◽  
Bicarbonate Transport ◽  
Key Words Cyanobacteria ◽  
Sodium Dependent ◽  
Rate Of Photosynthesis ◽  
Low Levels ◽  
In Cells

Cyanobacteria possess systems for the active transport of both CO2 and HCO3−. While the active CO2 transport system seems to be present in cells grown on all levels of CO2 or dissolved inorganic carbon, the bicarbonate transport systems are only present in cells grown on low levels of CO2 or dissolved inorganic carbon (air levels or lower). Active bicarbonate transport can be shown to occur when the rate of photosynthesis exceeds that which could be sustained by the production of CO2 from the dehydration of bicarbonate or when CO2 transport is inhibited with carbon oxysulfide or hydrogen sulfide. Two systems for active bicarbonate transport have been identified: one is dependent on the presence of millimolar concentrations of sodium, and the other is independent of the sodium requirement. Cells grown with air bubbling normally possess the first whereas cells grown in standing culture normally possess the second. The sodium-dependent bicarbonate transport can be inhibited by omitting sodium from the reaction medium or competitively with lithium when sodium is present. Monensin and amiloride also inhibit sodium-dependent bicarbonate transport. It does not appear to be inhibited by ethoxyzolamide. The inhibition of sodium-independent bicarbonate transport is not yet established. Bicarbonate transport appears to have no effect on CO2 transport and CO2 transport appears to have no effect on bicarbonate transport. Hence, the transport systems seems to be independent. Although a number of mechanisms have been proposed for bicarbonate transport, the experimental data are not sufficient to clearly distinguish between them. Key words: cyanobacteria, active CO2 transport, active HCO3− transport, photosynthesis, sodium.

Physiological aspects of CO2 and HCO3− transport by cyanobacteria: a review

1990 ◽  
Vol 68 (6) ◽  
pp. 1291-1302 ◽  
Author(s):  
Anthony G. Miller ◽  
George S. Espie ◽  
David T. Canvin
Keyword(s):  
Active Transport ◽  
Inorganic Carbon ◽  
Dissolved Inorganic Carbon ◽  
Structural Analog ◽  
Transport Systems ◽  
Ribulose Bisphosphate Carboxylase ◽  
High Concentration ◽  
Bisphosphate Carboxylase ◽  
Ribulose Bisphosphate Carboxylase Oxygenase ◽  
Inorganic Carbon Transport

Cyanobacteria grown at air levels of CO2, or lower, have a very high photosynthetic affinity for CO2. For ceils grown in carbon-limited chemostats at pH 9.6, the K0.5 (CO2) for whole cell CO2 fixation is about 3 nM. This is in spite of a K0.5 (CO2) for cyanobacterial ribulose bisphosphate carboxylase/oxygenase of about 200 μM. It is now clear that cyanobacteria can photosynthesize at very low CO2 concentrations because they raise the CO2 concentration dramatically around the carboxylase. This rise in the intracellular CO2 concentration involves the active transport of HCO3− and CO2, perhaps by separate transport systems. The transport of HCO3− often requires millimolar levels of Na+, and this provides a ready means of initiating HCO3− transport. The active transport of CO2 requires only micromolar levels of Na+. In the rather dense cell suspensions used in transport studies the extent of CO2 uptake is often limited by the rate at which CO2 can be formed from the HCO3− in the medium. The addition of carbonic anhydrase relieves this kinetic limitation on CO2 transport. The active transport of CO2 can be selectively inhibited by the structural analog carbon oxysulfide (COS). When HCO3− transport is allowed in the presence of COS there is a substantial net leakage of CO2 from the cells. This leaked CO2 results from the intracellular dehydration of the accumulated HCO3−. This CO2 is normally scavenged by the active CO2 pump. If cells are allowed to transport H13C18O18O18O− for 5 s and if CO2 transport is suddenly quenched by the addition of COS, then a rapid leakage of 13C16O16O occurs. If the rapidly released CO2 was actually present in the cells before the addition of the COS, then the intracellular CO2 concentration would have been about 0.6 mM. Not only is this a high concentration, but since the leaked CO2 was completely depleted of the initial 18O, it must have been in rapid equilibrium with the total dissolved inorganic carbon within the cells. Cells grown on high levels of inorganic carbon, either as CO2 or HCO3−, lack the active HCO3− system but still retain a capacity, albeit reduced, for CO2 transport. Cyanobacteria seem to adjust their complement of inorganic carbon transport systems so that the K0.5 for transport is close to the inorganic carbon concentration of the growth medium.

A physiological evaluation of carbon sources for calcification in the octocoral Leptogorgia virgulata (Lamarck).

1997 ◽  
Vol 200 (20) ◽  
pp. 2653-2662
Author(s):  
J M Lucas ◽  
L W Knapp
Keyword(s):  
Calcium Carbonate ◽  
Carbonic Anhydrase ◽  
Inorganic Carbon ◽  
Sea Water ◽  
Dissolved Inorganic Carbon ◽  
Carbon Sources ◽  
Iodoacetic Acid ◽  
Transport Systems ◽  
Disulfonic Acid ◽  
Leptogorgia Virgulata

The union of calcium cations with carbonate anions to form calcium carbonate (CaCO3) is a fundamentally important physiological process of many marine invertebrates, in particular the corals. In an effort to understand the sources and processes of carbon uptake and subsequent deposition as calcium carbonate, a series of studies of the incorporation of 14C-labeled compounds into spicules was undertaken using the soft coral Leptogorgia virgulata. It has been surmised for some time that dissolved inorganic carbon in sea water is used in the biomineralization process. Furthermore, it was suspected that metabolically generated CO2 is also available for calcification. As a means of testing these possible sources of carbon in spicule calcification, key enzymes or transport systems in each pathway were inhibited. First, the enzyme carbonic anhydrase was specifically inhibited using acetazolamide. Second, the active transport of bicarbonate was inhibited using DIDS (4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid). Third, CO2 generation resulting from glycolysis and the citric acid cycle was arrested using iodoacetic acid, which interferes specifically with the enzyme glyceraldehyde-3-phosphate dehydrogenase. The results indicate that dissolved CO2 is the largest source of carbon used in the formation of calcitic sclerites, followed by HCO3- from dissolved inorganic carbon. In L. virgulata, the dissolved inorganic carbon is responsible for approximately 67% of the carbon in the sclerites. The other 33% comes from CO2 generated by glycolysis. Two important conclusions can be drawn from this work. First, carbon for spiculogenesis comes not only from dissolved inorganic carbon in the environment but also from metabolically produced carbon dioxide. While the latter has been theorized, it has never before been demonstrated in octocorals. Second, regardless of the carbon source, the enzyme carbonic anhydrase plays a pivotal role in the physiology of spicule formation in Leptogorgia virgulata.

Comparative study of dissolved inorganic carbon utilization and photosynthetic responses in Nannochloris (Chlorophyceae) and Nannochloropsis (Eustigmatophyceae) species

1998 ◽  
Vol 76 (6) ◽  
pp. 1104-1108 ◽  
Author(s):  
I Emma Huertas ◽  
Luis M Lubián
Keyword(s):  
Life Cycle ◽  
Inorganic Carbon ◽  
Dissolved Inorganic Carbon ◽  
Nannochloropsis Oculata ◽  
Marine Phytoplankton ◽  
Bicarbonate Transport ◽  
Photosynthetic Oxygen Evolution ◽  
Carbon Utilization ◽  
Nannochloropsis Gaditana ◽  
Inorganic Carbon Utilization

Four species of marine microalgae with similar morphology and life cycle, namely Nannochloris atomus Butcher, Nannochloris maculata Butcher, Nannochloropsis gaditana Lubian, and Nannochloropsis oculata (Droop) Hibberd, have been examined with respect to their affinity for different sources of dissolved inorganic carbon. External carbonic anhydrase activity was not found in any of these species, but the cell affinity for dissolved inorganic carbon (DIC) in Nannochloris species was affected by the inhibitor acetazolamide at a concentration of 400 µM. Measurement of photosynthetic rates and CO2 compensation points at different pH values showed that the Nannochloris species had a greater capacity for CO2 rather than HCO3- utilization. In contrast, the observed rates of photosynthetic oxygen evolution in Nannochloropsis species were greater than could be accounted for by the theoretical rate of CO2 supply from the spontaneous dehydration of bicarbonate in the external medium. This indicates that these algae were able to transport bicarbonate across the plasmalemma. Furthermore, the K0.5 (DIC) value at acidic pH showed that Nannochloropsis oculata could also use CO2 as an exogenous carbon source for photosynthesis. Although the species of marine phytoplankton used in this study possess similar morphological characteristics and life cycle, there exist many differences in the mode of inorganic carbon utilization between these microalgae.Key words: Nannochloris, Nannochloropsis, inorganic carbon utilization, bicarbonate transport, CO2 compensation point, photosynthesis.

Regulation of inorganic carbon acquisition by phosphorus limitation in the green alga Chlorella emersonii

2005 ◽  
Vol 83 (7) ◽  
pp. 859-864 ◽  
Author(s):  
John Beardall ◽  
Simon Roberts ◽  
John A Raven
Keyword(s):  
Green Alga ◽  
Inorganic Carbon ◽  
Dissolved Inorganic Carbon ◽  
Stable Carbon Isotope ◽  
Phosphorus Limitation ◽  
Phosphate Limitation ◽  
P Limitation ◽  
Green Alga Chlorella ◽  
In Cells ◽  
Inorganic Carbon Acquisition

Inorganic phosphate (Pi) plays a central role in cellular energy transduction. As a consequence, limitation of growth by phosphate availability can have an important impact on various aspects of metabolism. Since carbon acquisition via CO2-concentrating mechanisms (CCMs) in most microalgae is an active process, requiring ATP, it might be expected that phosphate limitation could have an indirect regulatory influence on CCM activity. We grew the green alga Chlorella emersonii Shihira et Krauss in semicontinuous or continuous cultures in nutrient-replete conditions or with orthophosphate as the limiting nutrient. CCM activity was down-regulated by P limitation. K0.5(dissolved inorganic carbon) values increased from approximately 4.5 µmol·L–1 in cells growing at close to maximal rates to >12 µmol·L–1 in cells growing at 0.2 d–1. Maximal rates of photosynthesis decreased by approximately half over the same range of growth rates. Direct measurements of CCM activity showed that internal CO2 : external CO2 ratio was markedly decreased under P limitation, and concurrent measurements of stable carbon isotope discrimination were consistent with decreased CCM activity in the P-limited cells.Key words: phosphate, CCM, CO2-concentrating mechanism, Chlorella, inorganic carbon acquisition.

Oxygen photoreduction and its effect on CO2 accumulation and assimilation in air-grown cells of Synechococcus UTEX 625

1997 ◽  
Vol 75 (2) ◽  
pp. 274-283 ◽  
Author(s):  
Qinglin Li ◽  
David Thomas Canvin
Keyword(s):  
Light Intensity ◽  
Inorganic Carbon ◽  
Dissolved Inorganic Carbon ◽  
Reducing Power ◽  
High Light Intensity ◽  
High Light ◽  
Anaerobic Conditions ◽  
Key Words Cyanobacteria ◽  
Spectrometric Measurements ◽  
Oxygen Photoreduction

Mass spectrometric measurements of 16O2, 18O2, and 13CO2 were used to measure the rates of gross O2 evolution, O2 uptake, and CO2 assimilation in relation to light intensity, temperature, pH, and O2 concentration by air-grown cells of the cyanobacterium Synechococcus UTEX 625. CO2 fixation and O2 photoreduction increased with increased light intensity and, although CO2 fixation was saturated at 250 μmol ∙ m−2 ∙ s−1, O2 photoreduction was not saturated until about 550 μmol ∙ m−2 ∙ s−1. At high light intensity addition of inorganic carbon to the cells stimulated O2 photoreduction 2-fold when CO2, fixation was allowed and 5-fold when CO2, fixation was inhibited with iodoacetamide. The ability of O2, to act as an acceptor of photosynthetically generated reducing power was dependent upon the O2 concentration, and the substrate concentration required for half maximum rate (K½(O2)) was 53.2 ± 4.2 μM (mean ± SD, n = 3). The Q10 for oxygen photoreduction was about 2. A certain amount (10%) of O2 appeared to be required for maximum photosynthesis, as photosynthesis was inhibited under anaerobic conditions, especially at high light intensity. The point of inhibition is unknown but it seemed unlikely to be on CO2 transport or the concentration of intracellular dissolved inorganic carbon (Ci), as the rate of initial CO2 transport was enhanced and the intracellular Q1 pool increased in size under anaerobic conditions. Key words: cyanobacteria, photosynthesis, Ci concentrating mechanism, inorganic carbon pool, O2 photoreduction, electron transport, temperature.

scholarly journals Efficacy and Selectivity of FGF2-Saporin Cytosolically Delivered by PCI in Cells Overexpressing FGFR1

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1476
Author(s):  
Aurora K. Vikan ◽  
Michal Kostas ◽  
Ellen Margrethe Haugsten ◽  
Pål K. Selbo ◽  
Jørgen Wesche
Keyword(s):  
Treatment Options ◽  
Current Treatment ◽  
Fibroblast Growth Factor Receptors ◽  
Ribosome Inactivating Protein ◽  
Cytosolic Delivery ◽  
Low Levels ◽  
U2os Cells ◽  
Targeting Effect ◽  
Target Effects ◽  
In Cells

Fibroblast growth factor receptors (FGFRs) have become an attractive target in cancer research and therapy due to their implication in several cancers. Limitations of current treatment options require a need for additional, more specific and potent strategies to overcome cancers driven by FGFRs. Photochemical internalization (PCI) is a light-controlled method for cytosolic delivery of drugs that are entrapped in endosomes and lysosomes. We here evaluated the efficacy and selectivity of PCI of FGF2-saporin (FGF-SAP) in cells overexpressing FGFR1. FGF-SAP is a conjugate of FGF2 and the highly cytotoxic ribosome-inactivating protein (RIP) saporin, which is used as payload to eliminate cancer cells. Evaluation of the targeting effect of PCI of FGF-SAP was done by comparing the cytotoxic response in osteosarcoma cells with very low levels of FGFR1 (U2OS) to cells overexpressing FGFR1 (U2OS-R1). We demonstrate that PCI greatly enhances cytotoxicity of the drug showing efficient cell killing at pM concentrations of the drug in U2OS-R1 cells. However, U2OS cells were also sensitive to the toxin after PCI. Binding experiments using confocal microscopy and Western blotting techniques indicate that FGF-SAP is taken up by cells through heparan sulfate proteoglycans (HSPGs) in U2OS cells. We further show that the cytotoxicity of FGF-SAP in U2OS cells was reduced when cells were co-treated with heparin to compete out binding to HSPG, demonstrating that the cytotoxic effect was due to internalization by HSPGs. We conclude that to prevent off-target effects of FGF-based toxins, it will be necessary to circumvent binding to HSPGs, for example by mutating the binding site of FGF2 to HSPGs.

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