当前位置：
X-MOL 学术
›
Anal. Chem.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Redesigning Solvatochromic Probe Laurdan for Imaging Lipid Order Selectively in Cell Plasma Membranes
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-10-12 , DOI: 10.1021/acs.analchem.0c03559 Dmytro I. Danylchuk 1 , Erdinc Sezgin 2, 3 , Philippe Chabert 1 , Andrey S. Klymchenko 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-10-12 , DOI: 10.1021/acs.analchem.0c03559 Dmytro I. Danylchuk 1 , Erdinc Sezgin 2, 3 , Philippe Chabert 1 , Andrey S. Klymchenko 1
Affiliation
|
Imaging of biological membranes by environmentally sensitive solvatochromic probes, such as Laurdan, provides information about the organization of lipids, their ordering, and their uneven distribution. To address a key drawback of Laurdan linked to its rapid internalization and subsequent labeling of internal membranes, we redesigned it by introducing a membrane anchor group based on negatively charged sulfonate and dodecyl chain. The obtained probe, Pro12A, stains exclusively the outer leaflet of lipid bilayers of liposomes, as evidenced by leaflet-specific fluorescence quenching with a viologen derivative, and shows higher fluorescence brightness than Laurdan. Pro12A also exhibits stronger spectral change between liquid-ordered and liquid-disordered phases in model membranes and distinguishes better lipid domains in giant plasma membrane vesicles (GPMVs) than Laurdan. In live cells, it stains exclusively the cell plasma membranes, in contrast to Laurdan and its carboxylate analogue C-Laurdan. Owing to its outer leaflet binding, Pro12A is much more sensitive to cholesterol extraction than Laurdan, which is redistributed within both plasma membrane leaflets and intracellular membranes. Finally, its operating range in the blue spectral region ensures the absence of crosstalk with a number of orange/red fluorescent proteins and dyes. Thus, Pro12A will enable accurate multicolor imaging of lipid organization of cell plasma membranes in the presence of fluorescently tagged proteins of interest, which will open new opportunities in biomembrane research.
中文翻译：
重新设计Solvatochromic探针Laurdan，以选择性地成像细胞血浆膜中的脂质顺序。
通过对环境敏感的溶剂变色探针（例如Laurdan）对生物膜进行成像，可提供有关脂质组织，其有序性及其不均匀分布的信息。为了解决Laurdan与其快速内在化和随后内膜标记有关的关键缺陷，我们通过引入基于带负电荷的磺酸盐和十二烷基链的膜锚定基团来重新设计Laurdan。所获得的探针Pro12A仅染色脂质体脂质双层的外部小叶，如用紫精衍生物对小叶进行特异性荧光猝灭所证明，并且显示出比Laurdan更高的荧光亮度。Pro12A还在模型膜中的液相有序相和液相无序相之间表现出更强的光谱变化，并且与Laurdan相比，在巨质膜囊泡（GPMV）中区分出更好的脂质结构域。与Laurdan及其羧酸酯类似物C-Laurdan相反，在活细胞中，它仅染色细胞质膜。由于其外部小叶结合，Pro12A对胆固醇提取的敏感性比Laurdan高得多，Laurdan则在血浆膜小叶和细胞内膜中重新分布。最后，其在蓝色光谱区域内的工作范围可确保与许多橙色/红色荧光蛋白和染料不存在串扰。因此，在存在目标荧光标记的蛋白质的情况下，Pro12A将能够对细胞质膜的脂质组织进行准确的多色成像，
更新日期：2020-11-03
中文翻译：
重新设计Solvatochromic探针Laurdan，以选择性地成像细胞血浆膜中的脂质顺序。
通过对环境敏感的溶剂变色探针（例如Laurdan）对生物膜进行成像，可提供有关脂质组织，其有序性及其不均匀分布的信息。为了解决Laurdan与其快速内在化和随后内膜标记有关的关键缺陷，我们通过引入基于带负电荷的磺酸盐和十二烷基链的膜锚定基团来重新设计Laurdan。所获得的探针Pro12A仅染色脂质体脂质双层的外部小叶，如用紫精衍生物对小叶进行特异性荧光猝灭所证明，并且显示出比Laurdan更高的荧光亮度。Pro12A还在模型膜中的液相有序相和液相无序相之间表现出更强的光谱变化，并且与Laurdan相比，在巨质膜囊泡（GPMV）中区分出更好的脂质结构域。与Laurdan及其羧酸酯类似物C-Laurdan相反，在活细胞中，它仅染色细胞质膜。由于其外部小叶结合，Pro12A对胆固醇提取的敏感性比Laurdan高得多，Laurdan则在血浆膜小叶和细胞内膜中重新分布。最后，其在蓝色光谱区域内的工作范围可确保与许多橙色/红色荧光蛋白和染料不存在串扰。因此，在存在目标荧光标记的蛋白质的情况下，Pro12A将能够对细胞质膜的脂质组织进行准确的多色成像，
京公网安备 11010802027423号