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Membrane staining and phospholipid tracking in Pseudomonas aeruginosa PAO1 using the phosphatidylcholine mimic propargyl-choline
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Graham, Chris L. B., Bryant, Jack, Roper, David I. and Banzhaf, Manuel (2023) Membrane staining and phospholipid tracking in Pseudomonas aeruginosa PAO1 using the phosphatidylcholine mimic propargyl-choline. Access Microbiology . doi:10.1099/acmi.0.000690.v1 ISSN 2516-8290. (In Press)
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Official URL: http://doi.org/10.1099/acmi.0.000690.v1
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.
Item Type: | Journal Article | ||||||||||||||||||
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Subjects: | Q Science > QH Natural history Q Science > QP Physiology Q Science > QR Microbiology |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||||||||||||||
Library of Congress Subject Headings (LCSH): | Pseudomonas aeruginosa , Membrane lipids, Lecithin , Fluorescence spectroscopy | ||||||||||||||||||
Journal or Publication Title: | Access Microbiology | ||||||||||||||||||
Publisher: | Microbiology Society | ||||||||||||||||||
ISSN: | 2516-8290 | ||||||||||||||||||
Book Title: | Membrane staining and phospholipid tracking in Pseudomonas aeruginosa PAO1 using the phosphatidylcholine mimic propargyl-choline | ||||||||||||||||||
Official Date: | 16 August 2023 | ||||||||||||||||||
Dates: |
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DOI: | 10.1099/acmi.0.000690.v1 | ||||||||||||||||||
Status: | Peer Reviewed | ||||||||||||||||||
Publication Status: | In Press | ||||||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||||||||||||
Date of first compliant deposit: | 20 March 2024 | ||||||||||||||||||
Date of first compliant Open Access: | 20 March 2024 | ||||||||||||||||||
RIOXX Funder/Project Grant: |
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