Replacing plasma membrane outer leaflet lipids with exogenous lipid without damaging membrane integrity

• Published in: PloS one Vol. 14; no. 10; p. e0223572
• Main Authors: Li, Guangtao, Kakuda, Shinako, Suresh, Pavana, Canals, Daniel, Salamone, Silvia, London, Erwin
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 07.10.2019; Public Library of Science (PLoS)
• Subjects: Animals; beta-Cyclodextrins - pharmacology; Biochemistry; Biology; Biology and Life Sciences; Blood; Blood lipids; Cell Line, Tumor; Cell Membrane - drug effects; Cell Membrane - metabolism; Cell membranes; Cells (Biology); CHO Cells; Cholesterol; Composition; Cricetinae; Cricetulus; Damage prevention; EDTA; Exchanging; Humans; Lecithin; Lipid composition; Lipid Droplets - metabolism; Lipids; Localization; Mammalian cells; Mass spectrometry; Membrane lipids; Membrane structure; Membrane structures; Membrane vesicles; Methods; Penicillin; Phosphatidylcholine; Phosphatidylcholines - metabolism; Phospholipids; Plant lipids; Rabbits; Research and analysis methods; Scientific imaging; Sphingolipids; Sphingomyelin; Sphingomyelins - metabolism; Structure-function relationships; United States > US; Colorado
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0223572

We recently introduced a MαCD-based method to efficiently replace virtually the entire population of plasma membrane outer leaflet phospholipids and sphingolipids of cultured mammalian cells with exogenous lipids (Li et al, (2016) Proc. Natl. Acad. Sci USA 113:14025-14030). Here, we show if the lipid-to- MαCD ratio is too high or low, cells can round up and develop membrane leakiness. We found that this cell damage can be reversed/prevented if cells are allowed to recover from the exchange step by incubation in complete growth medium. After exchange and transfer to complete growth medium cell growth was similar to that of untreated cells. In some cases, cell damage was also prevented by carrying out exchange at close to room temperature (rather than at 37°C). Exchange with lipids that do (sphingomyelin) or do not (unsaturated phosphatidylcholine) support a high level of membrane order in lipid vesicles had the analogous effect on plasma membrane order, confirming exogenous lipid localization in the plasma membrane. Importantly, changes in lipid composition and plasma membrane properties after exchange and recovery persisted for several hours. Thus, it should be possible to use lipid exchange to investigate the effect of plasma membrane lipid composition upon several aspects of membrane structure and function.