Lipid Biology Laboratory, RIKEN, 2-1, Hirosawa, Wako-shi, Saitama, 351-0198, Japan.
Graduate School of Humanities and Life Sciences, Tokyo Kasei University, 1-18-1, Kaga, Itabashi, Tokyo, 173-8602, Japan.
Sci Rep. 2017 Jun 6;7(1):2900. doi: 10.1038/s41598-017-02980-x.
We have developed and characterized a novel photoswitchable phospholipid analog termed N-nitroBIPS-DPPG. The fluorescence can be switched on and off repeatedly with minimal photobleaching by UV or visible light exposure, respectively. The rather large photochromic head group is inserted deeply into the interfacial membrane region conferring a conical overall lipid shape, preference for a positive curvature and only minimal intermembrane transfer. Utilizing the switchable NBD fluorescence quenching ability of N-nitroBIPS-DPPG, a detergent free intermembrane transfer assay system for NBD modified lipids was demonstrated and validated. As NBD quenching can be turned off, total NBD associated sample fluorescence can be determined without the need of detergents. This not only reduces detergent associated systematic errors, but also simplifies assay handling and allows assay extension to detergent insoluble lipid species.
我们开发并表征了一种新型的可光开关的磷脂类似物，称为 N-硝基 BIPS-DPPG。通过分别用紫外光或可见光照射，可以反复打开和关闭荧光，且几乎没有光漂白。较大的光致变色头部基团深入插入界面膜区域，赋予了锥形的整体脂质形状，有利于正曲率，并且仅发生最小程度的膜间转移。利用 N-硝基 BIPS-DPPG 的可开关 NBD 荧光猝灭能力，展示并验证了用于 NBD 修饰脂质的无去污剂的膜间转移测定系统。由于可以关闭 NBD 猝灭，因此无需去污剂即可确定与总 NBD 相关的样品荧光。这不仅减少了与去污剂相关的系统误差，而且简化了测定操作，并允许测定扩展至不溶于去污剂的脂质种类。
