The mechanism of calcium uptake by liver microsomes: effect of anions and ionophores

Abstract

The mechanism of calcium uptake by liver microsomes was investigated using various anions and ionophores. Calcium uptake was shown to be specific to microsomes and unlikely to be due to contamination by plasma membranes by correlation of calcium uptake to the marker enzymes specific for these two fractions. Under the conditions employed, phosphates, sulfate, chloride, acetate, nitrate, thiocyanate, maleate, succinate and oxalate all stimulated calcium uptake by microsomes, but to different degrees. The greatest effect (4–6-fold) was observed with phosphate. On the contrary, phosphate is the only anion that stimulates the plasma membrane calcium uptake to any significant degree. Treatment of isolated microsomes with 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene (DIDS) resulted in inhibition of ATP- and anion-dependent calcium uptake. A lipid-permeable organic acid such as maleate retained its ability to promote calcium uptake in DIDS-treated microsomes. However, a lipophilic anion, such as nitrate, stimulated calcium uptake only in the presence of the protonophore carbonyl cyanide m- chlorophenylhydrazone (CCCP). In addition, 2 μM valinomycin, when added in the absence or presence of 10 to 100 mM K +, had no stimulatory effect on caicium uptake. These resuits appear to be consistent with a model in which the active uptake of calcium into microsomes involves electroneutral Ca 2+-n H + exchange.