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J. Biochem, 2003, Vol. 134, No. 1 121-128
© 2003 Japanese Biochemical Society

BIOTECHNOLOGY

Susceptibilities of Phospholipid Vesicles Containing Different Sterols to Amphotericin B–Loaded Lysophosphatidylcholine Micelles

Maki Onda, Yukari Inoue, Manami Kawabata and Tomoyoshi Mita+

Department of Environmental Sciences, Faculty of Science, Osaka Women’s University, 2-1 Daisen-cho, Sakai, Osaka 590-0035

To investigate the susceptibilities of fungal and mammalian cells to amphotericin B (AmB), AmB-loaded lysophosphatidylcholine (LPC)micelles as drug delivery vehicles were incubated at 37°C with phosphatidylcholine vesicles containing different sterols as model systems for fungal and mammalian cells. The binding and kinetics of AmB to sterols in the membranes were judged by UV-visible spectroscopy. In the 91% monomeric form, AmB interacted rapidly with ergosterol and slowly with 7-dehydrocholesterol (7-DHC), while it did not interact with cholesterol. In the 50% monomeric form, AmB formed complexes more rapidly with ergosterol or 7-DHC than in the monomeric form, whereas it did not still interact with cholesterol. The interaction was also characterized by resonance energy transfer between the fluorescent probe trimethylammonium diphenylhexatriene (TMA-DPH) and AmB. In the 91% monomeric form, AmB caused initial fluorescence quenching in bilayer membranes containing any sterol as well as sterol-free bilayer membranes due to the release of AmB and its incorporation within the membranes. However, a second phase of increasing fluorescence was found in the case of ergosterol alone. On the other hand, in the 47% monomeric form, AmB gave a biphasic intensity profile in membranes containing any sterol as well as sterol-free membranes. However, the extent of the second phase of increasing fluorescence intensity was markedly dependent upon sterol composition. Studies using sterol-containing vesicles provide important insights into the role of the aggregation state of AmB in its effects on cells.

+ To whom correspondence should be addressed. Fax: +81-722-22-4791, E-mail: mitaÅócenter.osaka-wu.ac.jp


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N. Yoshida, T. Mita, and M. Onda
Susceptibilities of Phospholipid Membranes Containing Cholesterol or Ergosterol to Gramicidin and its Derivative Incorporated in Lysophospholipid Micelles
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