Doxorubicin inhibits the phosphate-transport protein reconstituted in liposomes. A study on the mechanism of the inhibition.
The phosphate transport protein (PTP) has been isolated from beef heart mitochondria in the presence of cardiolipin and reconstituted in asolectin and phosphatidylcholine vesicles. As expected, the activity of the reconstituted PTP is inhibited by N-ethylmaleimide and mersalyl. It is also inhibited by adriamycin and Br-daunomycin. Storage of isolated mitochondria at -80 degrees C prior to the isolation of the PTP decreases the maximal activity of the unidirectional transport of phosphate (Pi) in reconstituted vesicles. The sensitivity of the system to the four inhibitors remains the same. The inhibition of the PTP by adriamycin is reversed by KCl and prevented by MgSO4. Since the interaction between adriamycin and cardiolipin is of ionic type, KCl apparently replaces adriamycin on the negative charges of cardiolipin. By contrast, MgSO4 complexes adriamycin directly, eliminating its inhibitory effect on the PTP. No PTP activity is found after reconstitution in phosphatidylcholine vesicles in the absence of cardiolipin. Addition of buffer-dispersed cardiolipin to the vesicles restores the Pi-transport activity. The addition of adriamycin to the vesicles together with cardiolipin removes the reactivation. Succinylation of the PTP at pH 8.0 eliminates the inhibitory effect of adriamycin in the reconstituted system. The effects of N-ethylmaleimide and mersalyl are not modified. The succinylated preparation reconstituted in phosphatidylcholine vesicles is reactivated by cardiolipin, but in this case the reactivation is not counteracted by adriamycin. Succinylation of the PTP at pH 9.0 results in its complete inhibition. The results indicate, (a) that the sites of interaction of MalNEt /mersalyl and adriamycin with the PTP X cardiolipin complex are different, and (b) that the adriamycin-binding site in the complex is probably on cardiolipin molecules essential to the PTP activity. It is postulated that succinylation of the PTP prevents the interaction of adriamycin with these molecules.
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002308:Cardiolipins / Q000494:pharmacology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002384:Catalysis; D002417:Cattle; D004317:Doxorubicin / Q000494:pharmacology*; D005033:Ethylmaleimide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000378:metabolism*; D008634:Mersalyl / Q000494:pharmacology; D008929:Mitochondria, Heart / Q000378:metabolism; D028044:Phosphate-Binding Proteins
D002308:Cardiolipins; D002352:Carrier Proteins; D008081:Liposomes; D028044:Phosphate-Binding Proteins; D008634:Mersalyl; D004317:Doxorubicin; D005033:Ethylmaleimide