Role of Photosynthetic Reactions in the Activity of Carbonic Anhydrase in Synechococcus sp. (UTEX 2380) in the Light : Inhibitor Studies Using the O-Exchange in C/O-Labeled Bicarbonate

Abstract

The role of the photosystems in the exchange of (18)O between species of inorganic carbon and water was studied in suspensions of the cyanobacterium Synechococcus sp. (UTEX 2380) using membrane-inlet mass spectrometry. This (18)O exchange is caused by the hydration-dehydration cycle of CO(2) and is catalyzed by carbonic anhydrase. We observed the complex (18)O exchange kinetics including dark-light-dark transients in suspensions of whole cells and found these to be identical to the (18)O 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 (18)O 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 (18)O exchange. Iodoacetamide, an inhibitor of CO(2) fixation, enhanced the (18)O 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 (18)O 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 CO(2) and HCO(3) (-) resulting in the (18)O exchange from inorganic carbon to water.