title: Effect of calmodulin antagonists on lysosomal enzyme secretion and phospholipid metabolism in guinea-pig macrophages.
abstract: The effects of calmodulin antagonists on the secretion of lysosomal enzyme and lipid metabolism in guinea-pig peritoneal macrophages were studied. Calmodulin antagonists, such as trifluoperazine, dibucaine and quinacrine, inhibited the secretion of N-acetyl-beta-d-glucosaminidase from cytochalasin B-treated macrophages when the macrophages were stimulated by the chemotactic peptide, formylmethionyl-leucyl-phenylalanine (f Met-Leu-Phe) or the Ca(2+) ionophore A23187. The effect of calmodulin antagonists on the incorporation of [(32)P]P(i) or [(3)H]glycerol into glycerolipids as well as on the redistribution of [(14)C]glycerol or [(3)H]arachidonic acid in [(14)C]glycerol- or [(3)H]arachidonic acid-prelabelled lipids were examined. Trifluoperazine, dibucaine or quinacrine stimulated [(32)P]P(i) incorporation into phosphatidic acid (PtdA) and phosphatidylinositol (PtdIns) without significant effect on the labelling of phosphatidylethanolamine (PtdEtn), phosphatidylserine (PtdSer), lysophosphatidylcholine (lyso-PtdCho) and lysophosphatidylethanolamine (lyso-PtdEtn). The incorporation of [(32)P]P(i) into phosphatidylcholine (PtdCho) was, on the contrary, inhibited. When calmodulin antagonists were added to macrophages stimulated by fMet-Leu-Phe, [(32)P]P(i) incorporation into PtdIns and PtdA was synergistically increased compared with that induced only by calmodulin antagonists. Trifluoperazine inhibited the incorporation of [(3)H]glycerol into PtdCho, triacylglycerol and PtdEtn. Also in this case, the incorporation of [(3)H]glycerol into PtdA and PtdIns was greatly enhanced. But [(3)H]glycerol incorporation into PtdSer, lyso-PtdEtn and lyso-PtdCho was not affected by the drug. On the other hand, diacylglycerol labelling with [(3)H]glycerol was maximally activated by 10mum-trifluoperazine and levelled off with the increasing concentration. When the effect of calmodulin antagonists on the redistribution of [(14)C]glycerol among lipids was examined in pulse-chase experiments, no significant effect on [(14)C]glycerol redistribution in PtdEtn, PtdCho, PtdIns, PtdSer, PtdA and tri- and di-acylglycerol could be detected. When macrophages prelabelled with [(3)H]arachidonic acid were treated with trifluoperazine, dibucaine or quinacrine, the [(3)H]arachidonic acid moiety in PtdEtn and PtdCho was decreased and that in PtdA was increased. The formation of [arachidonate-(3)H]diacylglycerol and non-esterified [(3)H]-arachidonic acid was also enhanced, but the increase in [(3)H]arachidonic acid was only observed at concentrations between 1 and 50mum. [Arachidonate-(3)H]PtdIns was not significantly affected. The activated formation of [arachidonate-(3)H]PtdA, diacylglycerol and non-esterified arachidonic acid by these drugs was synergistically enhanced in the presence of fMet-Leu-Phe.
mesh_terms: D000118:Acetylglucosaminidase / Q000378:metabolism*; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000037:antagonists & inhibitors*; D002147:Calmodulin / Q000037:antagonists & inhibitors*; D003992:Dibucaine / Q000494:pharmacology; D005260:Female; D006168:Guinea Pigs; D006596:Hexosaminidases / Q000378:metabolism*; D066298:In Vitro Techniques; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology*; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D011796:Quinacrine / Q000494:pharmacology; D014268:Trifluoperazine / Q000494:pharmacology; D014280:Triglycerides / Q000378:metabolism*
chemical_list: D001095:Arachidonic Acids; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D010743:Phospholipids; D014280:Triglycerides; D014268:Trifluoperazine; D016718:Arachidonic Acid; D006596:Hexosaminidases; D000118:Acetylglucosaminidase; D011796:Quinacrine; D003992:Dibucaine