Abstract
A mammalian plasma membrane is a structure on which several layers of complexity are built. The first order of complexity comes from the heterogeneity of lipid-ordered domains. Gangliosides in concert with cholesterol are preferentially packed on the outer leaflet and form lipid-ordered domains, commonly known as lipid rafts. The formation and dynamics of these domains impact nearly all membrane protein functions and are an intensely studied topic. However, tools suited for lipid domain alteration are extremely limited. Currently, methyl-β-cyclodextrin (MβCD) appears to be the most common way to disrupt lipid domains, which is believed to operate via cholesterol extraction. This significantly limits our ability in membrane biophysics research. Previously, we found that N-(3-oxo-dodecanoyl) homoserine lactone (3oc), a small signaling chemical produced by Pseudomonas aeruginosa, is highly efficient in altering lipid-ordered domains. In this study, 3oc was compared with MβCD in a series of biochemical, biophysical, and cell biological analyses. Per molarity, 3oc is more efficient than MβCD in domain alteration and appears to better retain membrane lipids after treatment. This finding will provide an essential reagent in membrane biophysics research.
Highlights
The eukaryotic plasma membrane is heterogeneous in lipid composition as well as in protein contents
We found that the LasI-LasR circuit of quorum sensing in Pseudomonas aeruginosa relies on the production of N-(3-oxo-dodecanoyl) homoserine lactone (3oc) (Song et al, 2019). 3oc is essential for P. aeruginosa to inhibit host immune attack
Our results suggest that 3oc is a viable replacement for MβCD for the purpose of lipid-ordered domain disruption
Summary
The eukaryotic plasma membrane is heterogeneous in lipid composition as well as in protein contents. Coupled to the repulsion of polyunsaturated phospholipids, the cone-shaped gangliosides are stabilized by small cholesterol to form a vertically slightly elevated area known as lipid-ordered domain (Lo, known as lipid rafts) in reference with the rest of the membrane (Ld or lipid disordered domains) (García-Arribas et al, 2016; Wang et al, 2017). These domains are highly dynamic and are in constant transition of formation (demixing) and dissolution (mixing) (Kusumi et al, 2005). Results indicate that the 3oc is comparable to MβCD by several key parameters and with some beneficial characters and suits the needs for lipid domain research
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