Activation of phospholipase A2 by adriamycin in vitro. Role of drug-lipid interactions.
To probe adriamycin-phospholipid interactions, the effects of this cytotoxin on the hydrolysis of a pyrene-labeled acidic alkyl-acyl phospholipid analog 1-octa-cosanyl-2-(6-pyren-1-yl)hexanoyl-sn-glycero-3-phos p hatidylmethanol (C28-O-PHPM) by porcine pancreatic phospholipase A2 (PLA2) were studied. In the absence of added Ca2+ adriamycin caused a 3-4-fold activation of hydrolysis of this pyrenelipid whereas an inhibition of action of PLA2 on the corresponding phosphatidylcholine derivative C28-O-PHPC was observed. Under similar conditions adriamycin also enhanced the rate of hydrolysis of the pyrene-labeled diacyl lipid 1-palmitoyl-2-(pyren-1-yl)hexanoyl-sn-glycero-3-phosphatidylgly cer ol and inhibited the hydrolysis of PLA2 on the phosphatidylcholine derivative. Increasing calcium concentrations abolished the activating and most of the inhibitory effects of adriamycin with the above phospholipid substrates. Quenching of pyrene excimer fluorescence by adriamycin revealed efficient binding of the drug to acidic lipids. Addition of 1 mM calcium reduced fluorescence quenching by adriamycin maximally by approximately 90%. In comparison, quenching by adriamycin of pyrene-labeled phosphatidylcholine was much weaker and calcium had only an insignificant effect. Monolayer experiments at an air/water interface showed a rapid and surface pressure-dependent penetration of the drug into a film of C28-O-PHPM. Increase in surface pressure was reversed by 80% by the inclusion of 1 mM Ca2+ into the subphase. Penetration of adriamycin into a monolayer of C28-O-PHPC was much weaker. In agreement with earlier studies two types of binding of adriamycin to C28-O-PHPM are proposed.
Mustonen
P
P
Department of Medical Chemistry, University of Helsinki, Finland.
Kinnunen
P K
PK
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004317:Doxorubicin / Q000378:metabolism / Q000494:pharmacology*; D004789:Enzyme Activation; D006868:Hydrolysis; D050356:Lipid Metabolism; D010179:Pancreas / Q000201:enzymology; D010713:Phosphatidylcholines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D013050:Spectrometry, Fluorescence; D013552:Swine
D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D004317:Doxorubicin; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium