Kinetic Analyses of Calcium Movements in HeLa Cell Cultures : II. Calcium efflux

Calcium efflux was studied in monolayers of HeLa cells. The fast phase of exchange was studied in an open system by continuous washout. Its half-time was 1.58 min which is practically identical to the fast phase of calcium influx previously found to be 1.54 min. This suggests that the fast component of efflux represents calcium exchange from an extracellular compartment probably from calcium bound to the cell membrane surface. Dinitrophenol (DNP) and iodoacetate (IAA) do not inhibit calcium efflux from this compartment. The slow phase of calcium exchange was studied in a closed three compartment system. The half-time of calcium efflux measured under these conditions is almost identical to that obtained previously in studies of calcium influx: 33.0 and 37.0 min, respectively. This slow compartment is likely to be the intracellular exchangeable calcium pool. DNP and IAA inhibit calcium efflux from this compartment, lengthening the half-time from 33 min to 55.0 and 216 min, respectively. This suggests that calcium extrusion from the cell is an active process. Since calcium influx is not affected by metabolic inhibitors, the cellular calcium concentration increases as would be predicted under these conditions. Calcium efflux is also markedly depressed by lowering the temperature.