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Metabolism and Enzymology

Role of Photosynthetic Reactions in the Activity of Carbonic Anhydrase in Synechococcus sp. (UTEX 2380) in the Light ¹

Inhibitor Studies Using the ¹⁸O-Exchange in ¹³C/¹⁸O-Labeled Bicarbonate

Department of Pharmacology, University of Florida, Gainesville, Florida 32610

The role of the photosystems in the exchange of ¹⁸O between species of inorganic carbon and water was studied in suspensions of the cyanobacterium Synechococcus sp. (UTEX 2380) using membrane-inlet mass spectrometry. This ¹⁸O exchange is caused by the hydration-dehydration cycle of CO₂ and is catalyzed by carbonic anhydrase. We observed the complex ¹⁸O exchange kinetics including dark-light-dark transients in suspensions of whole cells and found these to be identical to the ¹⁸O exchange kinetics of physiologically fully active spheroplast preparations. There was no enhancement effect of inorganic nitrogen on inorganic carbon accumulation. Membrane preparations exhibited no uptake of inorganic carbon and very little carbonic anhydrase activity, although these membranes were photosynthetically fully competent. DCMU, the inhibitor of photosystem II, eliminated almost entirely the ¹⁸O exchange activity of whole cells in the light. But this effect of DCMU could be reversed by addition of the electron donor couple 3,6-diaminodurene/ascorbate, suggesting the involvement of photosystem I in the events leading to ¹⁸O exchange. Iodoacetamide, an inhibitor of CO₂ fixation, enhanced the ¹⁸O exchange in whole cell suspensions and inhibited neither the uptake of inorganic carbon nor the dehydration of bicarbonate in the light. The proton carrier carbonylcyanide m-chlorophenylhydrazone and the inhibitors diethylstilbestrol and N,N' -dicyclohexyl carbodiimide affecting the membrane potential, totally abolished ¹⁸O exchange in the light. From ¹⁸ O-labeled inorganic carbon experiments we conclude that one of the roles of photosystem I is to provide the active uptake of inorganic carbon into the cells, where carbonic anhydrase catalyzes the interconversion between CO₂ and HCO₃^– resulting in the ¹⁸O exchange from inorganic carbon to water.