RT Journal Article
SR Electronic
T1 Polarization-sensitive stimulated Raman scattering imaging resolves amphotericin B orientation in <em>Candida</em> membrane
JF Science Advances
JO Sci Adv
FD American Association for the Advancement of Science
SP eabd5230
DO 10.1126/sciadv.abd5230
VO 7
IS 2
A1 Dong, Pu-Ting
A1 Zong, Cheng
A1 Dagher, Zeina
A1 Hui, Jie
A1 Li, Junjie
A1 Zhan, Yuewei
A1 Zhang, Meng
A1 Mansour, Michael K.
A1 Cheng, Ji-Xin
YR 2021
UL http://advances.sciencemag.org/content/7/2/eabd5230.abstract
AB Ergosterol-targeting amphotericin B (AmB) is the first line of defense for life-threatening fungal infections. Two models have been proposed to illustrate AmB assembly in the cell membrane; one is the classical ion channel model in which AmB vertically forms transmembrane tunnel and the other is a recently proposed sterol sponge model where AmB is laterally adsorbed onto the membrane surface. To address this controversy, we use polarization-sensitive stimulated Raman scattering from fingerprint C═C stretching vibration to visualize AmB, ergosterol, and lipid in single fungal cells. Intracellular lipid droplet accumulation in response to AmB treatment is found. AmB is located in membrane and intracellular droplets. In the 16 strains studied, AmB residing inside cell membrane was highly ordered, and its orientation is primarily parallel to phospholipid acyl chains, supporting the ion channel model. Label-free imaging of AmB and chemical contents offers an analytical platform for developing low-toxicity, resistance-refractory antifungal agents.