Abstract：
Monitoring active membrane cholesterol and lipid raft cholesterol in the inner leaflet of the plasma membrane is significant for understanding the membrane function and cellular physiopathological processes. Limited by existing methods, it is difficult to differentiate active membrane cholesterol and lipid raft cholesterol. A novel dual-monomer solvatochromic probe system (DSPS) that targets two types of cholesterol was developed. Acrylodan-BG/SNAP-D4 composed of SNAP-D4 cholesterol-recognizing monomers and solvatochromic acrylodan-BG-sensing monomers exhibits excellent cholesterol detecting properties in terms of selectivity, accuracy, convenience and economic benefits. Cell imaging revealed that lipid raft cholesterol emitted blue fluorescence, whereas active membrane cholesterol (which partially bobbed in aqueous cytosol) displayed green fluorescence; both the fluorescence emissions increased or decreased in a cholesterol-dependent manner. This system provides a new technology for the determination of two types of cholesterol, which is beneficial for the further study of membrane function, intracellular cholesterol trafficking, and cell signaling.
摘要：
监测质膜内膜中的活性膜胆固醇和脂筏胆固醇对于了解膜功能和细胞生理病理过程具有重要意义。受现有方法的限制，很难区分活性膜胆固醇和脂筏胆固醇。开发了一种针对两种类型胆固醇的新型双单体溶剂化变色探针系统（DSPS）。由SNAP-D4胆固醇识别单体和溶剂化显色的丙烯酰氨-BG敏感单体组成的丙烯酰氨-BG/SNAP-D4在选择性方面表现出优异的胆固醇检测性能，准确度,方便和经济效益。细胞成像显示脂筏胆固醇发出蓝色荧光，而活性膜胆固醇（部分在细胞溶胶中）显示绿色荧光；两种荧光发射均以胆固醇依赖性方式增加或减少。该系统为两种胆固醇的测定提供了一种新的技术，有利于进一步研究膜的功能,细胞内胆固醇运输，和细胞信号。
