Membrane staining and phospholipid tracking in Pseudomonas aeruginosa PAO1 using the phosphatidylcholine mimic propargyl-choline
Membrane staining and phospholipid tracking in Pseudomonas aeruginosa PAO1 using the phosphatidylcholine mimic propargyl-choline
复制标题
使用模拟磷脂酰胆碱对铜绿假单胞菌 PAO1 进行膜染色和磷脂追踪
DOI：
10.1099/acmi.0.000690.v1
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Graham C
中科院分区：
文献类型：
--
作者：
Graham C
在活体荧光显微镜中使用膜特定的染料是很常见的。然而，这些试剂大多是非特异性的，不能跟踪特定的脂类物种的运动，而是经常作为非共价脂类相关的探针或需要将完整的脂类和酰尾摄取到膜中。这个问题已经在真核细胞生物学中通过使用易于点击化学的磷脂头基脉冲标记得到了解决。在这里，我们描述了一种在铜绿假单胞菌中通过荧光成像标记磷脂的方法，该方法使用一种磷脂酰胆碱模拟物‘丙叉胆碱’(PCho)。这种易于点击化学的头基模拟物可通过显微镜观察，并允许共价标记脂类。细胞膜的荧光在不同的斑块中可见，依赖于PCHO的浓度，并且定位于细胞膜部分，表明它适合于膜标记和细胞成像。
The use of membrane-specific dyes forin vivofluorescent microscopy is commonplace. However, most of these reagents are non-specific and cannot track specific lipid species movement, instead often acting as non-covalent lipid-associated probes or requiring the uptake of whole lipids and acyl tails into the membrane. This issue has been solved in eukaryotic cell biology by the use of click-chemistry-liable phospholipid headgroup pulse labels. Here, we describe a method forin vivophospholipid labelling by fluorescent imaging inPseudomonas aeruginosausing a phosphatidylcholine mimic, ‘propargyl-choline’ (PCho). This click-chemistry-liable headgroup mimic is visible by microscopy and allows the covalent labelling of lipids. Fluorescence of the cell membranes, visible in heterogeneous patches, is dependent on PCho concentration and is localized in the membrane fraction of cells, demonstrating that it is suitable for membrane labelling and cell imaging.