期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1863, 期 8, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.bbamem.2021.183626
关键词
Cholesterol; Completely miscible; Phase separation; Fluorescence microscope; Atomic force microscope
资金
- JST-CREST [JPMJCR14F3]
- JST-A-STEP
- JSPS KAKENHI [JP20H02690]
- Casio Science Promotion Foundation
- Amano Institute of Technology
The study reported the formation of microdomains in a ternary lipid bilayer composed of PE, PC, and Chol, which functioned as specific fusion sites for proteoliposomes. By investigating the composition and mechanism of these unique microdomains in supported lipid bilayers, it was found that different types of PE affected the area fraction of microdomains and that the microdomains were rich in polyunsaturated PE and exhibited L-d-like phase. The association of eggPC and Chol resulted in microdomain formation by segregating polyunsaturated PE and facilitating Chol interactions in the hydrophobic core.
Recently, we reported that a ternary lipid bilayer comprising phosphatidylethanolamine (PE), phosphatidylcholine (PC), which were both derived from chicken egg, and cholesterol (Chol) generates microdomains that function as specific fusion sites for proteoliposomes. Chol-induced microdomain formation in a completely miscible lipid bilayer is an exceptional phenomenon. Numerous studies have elucidated the formation of domains in liquid ordered (L-o) and liquid disordered (L-d) phases of ternary bilayers, which comprise two partially miscible lipids and Chol. Herein, we investigated the composition and mechanism of formation of these unique microdomains in supported lipid bilayers (SLBs) using a fluorescence microscope and an atomic force microscope (AFM). We prepared ternary SLBs using egg-derived PC (eggPC), Chol and three different types of PE: egg-derived PE, 1-palmitoyl-2-oleoyl-PE, and 1,2-didocosahexaenoyl-PE (diDHPE). Fluorescence microscopy observations revealed that fluid and continuous SLBs were formed at PE concentrations (C-PE) of >= 6 mol%. Fluorescence recovery after photobleaching measurement revealed that the microdomain was more fluid than the surrounding region that showed typical diffusion coefficient of the L-o phase. The microdomains were observed as depressions in the AFM topographies. Their area fraction (theta) increased with CPE, and diDHPE produced a significantly large theta among the three PEs. The microdomains in the PE+eggPC+Chol-SLBs were rich in polyunsaturated PE and were in the L-d-like phase. Associating eggPC and Chol caused polyunsaturated PE to segregate, resulting in a microdomain formation by conferring the umbrella effect on Chol, entropic effect of disordered acyl chains, and pi-pi interactions in the hydrophobic core.
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