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ATP-induced beta-glucuronidase release from undifferentiated HL-60 cells is dependent on Ca2+ ions
D E Packham1, L Jiang, A D Conigrave
1Department of Biochemistry, University of Sydney, NSW, Australia.
Abstract:
Extracellular ATP (0.1-100 microM) evoked a biphasic increase in cytosolic free Ca2+ ion concentration ([Ca2+]i) in fura-2 loaded undifferentiated HL-60 cells and activated the release of the lysosomal enzyme beta-glucuronidase. The EC50 for the elevation of [Ca2+]i was about 0.2 microM. However, the EC50 for the activation of beta-glucuronidase release was two orders of magnitude higher (about 20 microM). These data imply that the ATP-induced elevation in [Ca2+]i is not sufficient to provoke beta-glucuronidase release. The role of [Ca2+]i in ATP-induced beta-glucuronidase release was further investigated using Ca(2+)-free solutions and by loading HL-60 cells with the non-fluorescent Ca(2+)-chelator BAPTA-AM. Ca2+ ions from both intracellular and extracellular sources were necessary for the activation of beta-glucuronidase secretion by extracellular ATP. However, the internal store is the major source of Ca2+ ions for the response.