Role of anionic phospholipids in the interaction of doxorubicin and plasma membrane vesicles: drug binding and structural consequences in bacterial systems.
Anthracycline-membrane interactions play a role in the transport, the cytoplasmic distribution, and possibly also the activity of anthracyclines. Previous work on model membranes has shown that the widely-applied anticancer drug doxorubicin interacts specifically with anionic phospholipids [de Wolf, F. A., et al. (1991) Biochim. Biophys. Acta 106, 67-80]. We have now been able to investigate these interactions, and their selectivity for anionic phospholipids, directly in plasma membranes. Because of the recent availability of Escherichia coli mutants in which the anionic phospholipid content ranges from only 10% to as much as 100% of the total phospholipid content, we used this bacterium as a source of plasma membranes. We compared the interactions of the cationic anthracycline doxorubicin with (1) plasma membranes of different mutant strains, (2) total lipid extracts of these membranes, and (3) synthetic phospholipid mixtures in which a comparable fraction of the phospholipids was negatively charged. The results show that anionic phospholipids are important determinants of doxorubicin binding, not only in model membranes but also in plasma membrane systems. Only in plasma membranes with a very low anionic lipid content was the binding to the anionic phospholipid masked by other factors. Using an unsaturated fatty acid auxotroph grown on [11,11-2H2]oleic acid, it appeared from 2H-NMR data that doxorubicin induces a disordering of acyl chains in bacterial plasma membranes and their total lipid extracts. This indicates that the binding is not purely electrostatic but involves the insertion of drug molecules into the lipid matrix, probably due to hydrophobic interactions.
de Wolf
F A
FA
Institute of Molecular Biology and Medical Biotechnology, University of Utrecht, The Netherlands.
Staffhorst
R W
RW
Smits
H P
HP
Onwezen
M F
MF
de Kruijff
B
B
D001665:Binding Sites; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D004317:Doxorubicin / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D008563:Membrane Lipids / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism / Q000494:pharmacology*; D013104:Spheroplasts / Q000378:metabolism
D008563:Membrane Lipids; D010743:Phospholipids; D004317:Doxorubicin