CHLORINATED HYDROCARBON-CELL MEMBRANE INTERACTIONS STUDIED BY THE FLUORESCENCE QUENCHING OF CARBAZOLE-LABELED PHOSPHOLIPIDS - PROBE SYNTHESIS AND CHARACTERIZATION OF THE QUENCHING METHODOLOGY
CHLORINATED HYDROCARBON-CELL MEMBRANE INTERACTIONS STUDIED BY THE FLUORESCENCE QUENCHING OF CARBAZOLE-LABELED PHOSPHOLIPIDS - PROBE SYNTHESIS AND CHARACTERIZATION OF THE QUENCHING METHODOLOGY
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DOI：
10.1016/0009-3084(80)90008-0
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发表时间：
1980-01-01
影响因子：
3.4
通讯作者：
HOGEN, D
中科院分区：
文献类型：
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作者：
LAKOWICZ, JR;HOGEN, D
认识到需要更好地了解氯化烃杀虫剂与细胞膜的[毒性]相互作用，研究了这些氯化烃对膜结合荧光团的荧光猝灭的用途。对潜在荧光团的广泛调查确定咔唑的 N-烷基衍生物是特别合适的荧光团。这些衍生物的荧光发射被多种常用的氯化烃猝灭。这种猝灭是碰撞性的，不会导致明显的光分解。合成了四种结构不同的咔唑标记磷脂，并通过 270 MHz PMR 以及色谱和化学手段确认了它们的结构。每个标记磷脂的咔唑部分应位于脂质双层的不同深度。水溶性猝灭剂表明荧光团无法进入水相，无论其与磷脂的连接点如何。当掺入脂质双层时，这些咔唑标记的磷脂的荧光寿命揭示了荧光团和氯化烃之间的碰撞频率。因此，膜结合荧光团的猝灭可用于测量：双层中氯化烃的扩散速率、脂质-水分配系数和膜对氯化烃的最大结合能力。给出了所有这些测量的示例，并通过直接化学分析确认了荧光测定结果。
In recognition of the need to better understand the [toxic] interactions of the chlorinated hydrocarbon insecticides with cell membranes, the use of fluorescence quenching of membrane-bound fluorophores by these chlorinated hydrocarbons was investigated. An extensive survey of potential fluorophores identified the N-alkyl derivatives of carbazole as being especially suitable fluorophores. The fluorescence emission of these derivatives is quenched by a wide variety of commonly-used chlorinated hydrocarbons. This quenching is collisional and does not result in significant photodecomposition. Four structurally distinct carbazole-labeled phospholipids were synthesized, and their structures were confirmed by 270 MHz PMR and by chromatographic and chemical means. The carbazole moiety of each labeled phospholipid should be localized at a different depth in lipid bilayer. Water soluble quenchers indicate that the fluorophores are inaccessible to the aqueous phase, irrespective of their point of attachment to the phospholipids. When incorporated into lipid bilayers, the fluorescence lifetime of these carbazole-labeled phospholipids reveals the collisional frequency between the fluorophore and the chlorinated hydrocarbon. As a result, quenching of membrane-bound fluorophores may be used to measure: the diffusional rate of the chlorinated hydrocarbon in the bilayer, the lipid-water partition coefficient and the maximum binding capacity of the membrane for the chlorinated hydrocarbon. Examples of all these measurements are given and the fluorometric results are confirmed by direct chemical analysis.