Formation of Cell Membrane Component Domains in Artificial Lipid Bilayer

Abstract

The lipid bilayer environment around membrane proteins strongly affects their structure and functions. Here, we aimed to study the fusion of proteoliposomes (PLs) derived from cultured cells with an artificial lipid bilayer membrane and the distribution of the PL components after the fusion. PLs, which were extracted as a crude membrane fraction from Chinese hamster ovary (CHO) cells, formed isolated domains in a supported lipid bilayer (SLB), comprising phosphatidylcholine (PC), phosphatidylethanolamine (PE), and cholesterol (Chol), after the fusion. Observation with a fluorescence microscope and an atomic force microscope showed that the membrane fusion occurred selectively at microdomains in the PC + PE + Chol-SLB, and that almost all the components of the PL were retained in the domain. PLs derived from human embryonic kidney 293 (HEK) cells also formed isolated domains in the PC + PE + Chol-SLB, but their fusion kinetics was different from that of the CHO-PLs. We attempted to explain the mechanism of the PL-SLB fusion and the difference between CHO- and HEK-PLs, based on a kinetic model. The domains that contained the whole cell membrane components provided environments similar to that of natural cell membranes, and were thus effective for studying membrane proteins using artificial lipid bilayer membranes.

FTIR spectra signatures reveal different cellular effects of EGFR inhibitors on nonsmall cell lung cancer cells

“…Cellular lipid composition has been considered as cellular dynamic. Lipid is a building block of cell membrane which involved in many cellular processes such as cell signaling, cellular transportation leading to form lipid domain . Lipid accumulation has recently been reported that induced cell death via apoptosis and established TKIs resistance on NSCLC cell line and patient tissues .…”

Supported Lipid Bilayers of Escherichia coli Extracted Lipids and Their Calcium Dependence

“…When the substrate was incubated in the PLE-suspension with [Ca 2+ ] = 2 mM, a circular region with a uniform fluorescence intensity existed ( Figure 1B). The fluorescence intensity shows that single layer of SLB was formed in this region (Tero et al, 2017;Kakimoto et al, 2018). The surrounding gray region contained bright dots that had similar fluorescence intensity to the circular SLB region (Figure 1B, inset).…”

“…They are used to investigate the fundamental physicochemical properties and molecular distribution of lipid bilayers with atomic force microscopy (AFM) and fluorescence-microscope-based methods (El Kirat et al, 2010;Tero, 2012;Zhong and He, 2012). Morphology and physicochemical properties of lipid domains, and shape transformation of membranes have been studied in multicomponent SLBs (El Kirat et al, 2010;Goertz et al, 2011;Zhong and He, 2012;Kawakami et al, 2017;Sumitomo and Oshima, 2017;Tero et al, 2017;Kakimoto et al, 2018).…”

Soft X-ray absorption spectroscopy in the low-energy region explored using an argon gas window

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