Extracellular ATP induces the release of calcium from intracellular stores without the activation of protein kinase C in Swiss 3T6 mouse fibroblasts.
Exposure of Swiss 3T6 mouse fibroblasts to extracellular ATP stimulated the formation of inositol phosphates and mobilized intracellular calcium. The mobilization of intracellular calcium was verified by imaging of fura-2 fluorescence in individual cells and by monitoring the efflux of 45Ca2+ from preloaded cells. However, we found no activation of protein kinase C as measured by phosphorylation of an 80-kDa acidic protein and by transmodulation of the receptor for epidermal growth factor. A careful examination of the kinetics of the phosphorylation reaction (from 30 sec to 10 min) revealed no activation of protein kinase C by extracellular ATP at any time. The lack of activation of protein kinase C was demonstrated even when a concentration of ATP 10-fold higher than that required to give a strong Ca2+ signal was used. Extracellular ATP did not inhibit protein kinase C activation by fetal bovine serum, platelet-derived growth factor, or phorbol esters. The effects of ATP were also produced by UTP but not by ADP, AMP, or adenosine. These findings demonstrate that it is possible to induce the mobilization of intracellular calcium by an inositol phosphate-mediated pathway without the activation of protein kinase C.
Gonzalez
F A
FA
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, NY 14853.
Rozengurt
E
E
Heppel
L A
LA
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D004789:Enzyme Activation; D005347:Fibroblasts / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D010750:Phosphoproteins / Q000096:biosynthesis / Q000302:isolation & purification; D011493:Protein Kinase C / Q000378:metabolism*; D012265:Ribonucleotides / Q000494:pharmacology; D014544:Uridine Triphosphate / Q000494:pharmacology
D010750:Phosphoproteins; D012265:Ribonucleotides; D000255:Adenosine Triphosphate; D011493:Protein Kinase C; D002118:Calcium; D014544:Uridine Triphosphate