title: Phase diagram of ternary cholesterol/palmitoylsphingomyelin/palmitoyloleoyl-phosphatidylcholine mixtures: spin-label EPR study of lipid-raft formation.
abstract: For canonical lipid raft mixtures of cholesterol (chol), N-palmitoylsphingomyelin (PSM), and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), electron paramagnetic resonance (EPR) of spin-labeled phospholipids--which is insensitive to domain size--is used to determine the ternary phase diagram at 23°C. No phase boundaries are found for binary POPC/chol mixtures, nor for ternary mixtures with PSM content <24 mol %. EPR lineshapes indicate that conversion from the liquid-disordered (L(α)) to liquid-ordered (L(o)) phase occurs continuously in this region. Two-component EPR spectra and several tie lines attributable to coexistence of gel (L(β)) and fluid phases are found for ternary mixtures with low cholesterol or low POPC content. For PSM/POPC alone, coexistence of L(α) and L(β) phases occurs over the range 50-95.5 mol % PSM. A further tie line is found at 3 mol % chol with endpoints at 50 and ≥77 mol % PSM. For PSM/chol, L(β)-L(o) coexistence occurs over the range 10-38 mol % chol and further tie lines are found at 4.5 and 7 mol % POPC. Two-component EPR spectra indicative of fluid-fluid (L(α)-L(o)) phase separation are found for lipid compositions: 25%<PSM<65%, 5%<chol<30-35%, 65%>POPC>10%, and confirmed by nonlinear EPR. Tie lines are identified in the L(α)-L(o) coexistence region, indicating that the fluid domains are of sufficient size to obey the phase rule. The three-phase triangle is bounded approximately by the compositions 40 and 75 mol % PSM with 10 mol % chol, and 60 mol % PSM with 25 mol % chol. These studies define the compositions of raft-like L(o) phases for a minimal realistic biological lipid mixture.
authors: Centre of Photochemistry, Russian Academy of Sciences, Moscow, Russia.
authors: Irina V
authors: IV
authors: Ionova
authors: Vsevolod A
authors: VA
authors: Livshits
authors: Derek
authors: D
authors: Marsh
mesh_terms: D002784:Cholesterol / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D021962:Membrane Microdomains / Q000737:chemistry*; D017711:Nonlinear Dynamics; D010713:Phosphatidylcholines / Q000737:chemistry*; D013109:Sphingomyelins / Q000737:chemistry*; D013113:Spin Labels
chemical_list: D010713:Phosphatidylcholines; D013109:Sphingomyelins; D013113:Spin Labels; C033171:palmitoylsphingomyelin; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine