Membrane staining and phospholipid tracking in Pseudomonas aeruginosa PAO1 using the phosphatidylcholine mimic propargyl-choline

Full text loading... Abstract The use of membrane-specific dyes for in vivo fluorescent 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 uptake of whole lipids and acyl tails into the membrane. This issue has been solved in eukaryotic cell biology by use of click-chemistry liable phospholipid headgroup pulse-labels. Here we describe a method for in vivo phospholipid labelling by fluorescent imaging in Pseudomonas aeruginosa using a phosphatidylcholine (PC) 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 heterogenous patches, is dependent on PCho concentration and is localised in the membrane fraction of cells, demonstrating that it is suitable for membrane labelling and cell imaging. - Received: - Version Posted: Funding - Antibiotic Research UK (Award ANTRUK_SRG_05-2021) - Principal Award Recipient: Chris LB Graham - Natural Environment Research Council (Award NE/T014717/1) - Principal Award Recipient: Chris LB Graham - Biotechnology and Biological Sciences Research Council (Award BB/M01116X/1) - Principal Award Recipient: David I Roper