Photoswitchable phospholipid FRET acceptor: Detergent free intermembrane transfer assay of fluorescent lipid analogs.
Photoswitchable phospholipid FRET acceptor: Detergent free intermembrane transfer assay of fluorescent lipid analogs.
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DOI：
10.1038/s41598-017-02980-x
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发表时间：
2017-06-06
影响因子：
4.6
通讯作者：
Kobayashi T
中科院分区：
文献类型：
--
作者：
Sumi M;Makino A;Inaba T;Sako Y;Fujimori F;Greimel P;Kobayashi T
我们开发并表征了一种新型光开关磷脂类似物，称为 N-硝基BIPS-DPPG。荧光可以反复打开和关闭，分别通过紫外线或可见光照射而产生最小的光漂白。相当大的光致变色头部基团深深地插入界面膜区域，赋予圆锥形的整体脂质形状，优选正曲率和仅最小的膜间转移。利用 N-硝基BIPS-DPPG 的可切换 NBD 荧光猝灭能力，对 NBD 修饰脂质的无去垢剂膜间转移测定系统进行了论证和验证。由于可以关闭 NBD 猝灭，因此可以在不需要去垢剂的情况下测定总 NBD 相关样品荧光。这不仅减少了与去垢剂相关的系统误差，而且还简化了测定处理并允许测定扩展到去垢剂不溶性脂质种类。
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.