Kotler-Brajtburg J, Medoff G, Schlessinger D, Kobayashi G S
Antimicrob Agents Chemother. 1974 Dec;6(6):770-6. doi: 10.1128/AAC.6.6.770.
Based on the enhanced fluorescence of amphotericin B in acid solutions, a quantitative assay for this polyene antibiotic has been developed that is sensitive and linear in the range of 0.1 to 10.0 muM. The binding of amphotericin B to Saccharomyces cerevisiae was assayed under various conditions as the amount bound to cells in a dialysis chamber or after centrifugation. Two types of binding were defined: weak, reversible binding occurred at 0 C or higher temperatures and even in the presence of inhibitors of energy metabolism, whereas strong, irreversible binding did not occur at 0 C and was inhibited when energy metabolism was blocked. Only strong binding was correlated with cell killing. Weak binding probably involves the outer layer of the membrane; strong binding probably requires disruption of hydrophobic regions of the cell membrane.
基于两性霉素B在酸性溶液中荧光增强的特性，已开发出一种针对这种多烯抗生素的定量测定方法，该方法在0.1至10.0 μM范围内灵敏且呈线性。在各种条件下测定两性霉素B与酿酒酵母的结合情况，可通过透析室中与细胞结合的量或离心后与细胞结合的量来进行。定义了两种结合类型：弱的、可逆的结合在0℃或更高温度下发生，甚至在存在能量代谢抑制剂的情况下也会发生；而强的、不可逆的结合在0℃时不发生，并且当能量代谢被阻断时会受到抑制。只有强结合与细胞杀伤相关。弱结合可能涉及细胞膜的外层；强结合可能需要破坏细胞膜的疏水区域。
