An adenosine triphosphate-dependent calcium uptake pump in human neutrophil lysosomes

Abstract

Regulation of the cytosolic free calcium concentrations is important to neutrophil function. In these studies, an ATP-dependent calcium uptake pump has been identified in human neutrophil lysosomes. This energy-dependent Ca++ uptake pump has a high affinity for Ca++ (Michaelis constant [K(m)] Ca++=107 nM) and a maximum velocity (V(max)) of 5.3 pmol/mg of protein per min. ATP was the only nucleotide that supported Ca++ uptake by lysosomes. The K(m) for ATP was 177 μM. ATP-dependent Ca++ uptake by neutrophil lysosomes was temperature- and pH-sensitive with optimal Ca++ pump activity at 37° C and pH 7.0-7.5. Mg++ was also essential for ATP-dependent Ca++ uptake by lysosomes. Azide and antimycin A had no effect on the energy-dependent uptake of Ca++ by neutrophil lysosomes. The chemotactic peptide formyl-methionyl-leucyl-phenylalanine inhibited ATP-dependent Ca++ accumulation by isolated lysosomes. Butoxycarbonyl-phenylalanine-leucine-phenylalanine-leucine-phenylalanin e, a competitive antagonist of the chemotactic peptide, blocked this inhibitory effect. These studies demonstrate the presence of an ATP-dependent Ca++ uptake pump in human neutrophil lysosomes that functions at physiologic intracellular concentrations of Ca++, ATP, and H+ and may be important to regulating neutrophil function by modulating cytosolic Ca++.