Abstract
For over 50 years, amphotericin has remained the powerful but highly toxic last line of defense in treating life-threatening fungal infections in humans with minimal development of microbial resistance. Understanding how this small molecule kills yeast is thus critical for guiding development of derivatives with an improved therapeutic index and other resistance-refractory antimicrobial agents. In the widely accepted ion channel model for its mechanism of cytocidal action, amphotericin forms aggregates inside lipid bilayers that permeabilize and kill cells. In contrast, we report that amphotericin exists primarily in the form of large, extramembranous aggregates that kill yeast by extracting ergosterol from lipid bilayers. These findings reveal that extraction of a polyfunctional lipid underlies the resistance-refractory antimicrobial action of amphotericin and suggests a roadmap for separating its cytocidal and membrane-permeabilizing activities. This new mechanistic understanding is also guiding development of what are to our knowledge the first derivatives of amphotericin that kill yeast but not human cells.
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Acknowledgements
We gratefully acknowledge P.J. Hergenrother and E. Oldfield for helpful discussions, and we thank J.J. Lopez for preliminary spin diffusion SSNMR experiments. Portions of this work were supported by the US National Institutes of Health (NIH) (R01GM080436 and F30DK081272) and the University of Illinois at Urbana-Champaign (Centennial Scholar Award to C.M.R.). M.D.B. is a Howard Hughes Medical Institute Early Career Scientist. M.C.C. is an American Heart Association Predoctoral Fellow. T.M.A. is a Ruth L. Kirchstein NIH National Research Service Award Predoctoral Fellow. The Gonen lab is funded by the Howard Hughes Medical Institute.
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T.M.A., M.C.C., A.G.C., K.A.D., A.J.N., G.C., T.G., C.M.R. and M.D.B. designed research. T.M.A., N.M. and A.G.C. prepared [U-13C]AmB and [13C]Erg. T.M.A., M.C.C., A.G.C., G.S.H., A.J.N., G.C. and B.E.U. prepared samples for SSNMR. M.C.C., A.J.N., G.C., G.S.H., M.D.T. and C.M.R. acquired SSNMR data. A.G.C. and T.G. performed microscopy. K.A.D. performed cell-based assays. T.M.A., M.C.C., A.G.C., K.A.D., G.S.H., M.D.T., A.J.N., G.C., S.W., B.E.U., E.L.W., T.G., C.M.R. and M.D.B. analyzed data. T.M.A., M.C.C., A.G.C., K.A.D., C.M.R. and M.D.B. wrote the paper.
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Biosynthesis and characterization of U-13C-Amphotericin Biosynthesis (DOCX 45 kb)
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Anderson, T., Clay, M., Cioffi, A. et al. Amphotericin forms an extramembranous and fungicidal sterol sponge. Nat Chem Biol 10, 400–406 (2014). https://doi.org/10.1038/nchembio.1496
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DOI: https://doi.org/10.1038/nchembio.1496
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