Lipid−Protein Interactions and Effect of Local Anesthetics in Acetylcholine Receptor-Rich Membranes from Torpedo marmorata Electric Organ

• Published in: Biochemistry (Easton) Vol. 42; no. 30; pp. 9167 - 9175
• Main Authors: Mantipragada, S. B, Horváth, L. I, Arias, H. R, Schwarzmann, G, Sandhoff, K, Barrantes, F. J, Marsh, D
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 05.08.2003
• Subjects: Anesthetics, Local - pharmacology; Animals; Benzocaine - pharmacology; Binding, Competitive - drug effects; Electric Organ - drug effects; Electric Organ - metabolism; Electron Spin Resonance Spectroscopy; Gangliosides - metabolism; Hydrophobic and Hydrophilic Interactions; Membrane Proteins - metabolism; Models, Chemical; Phosphatidylinositols - metabolism; Phospholipids - metabolism; Procaine - pharmacology; Receptors, Nicotinic - metabolism; Spectrometry, Fluorescence; Spin Labels; Stearic Acids - metabolism; Synaptic Membranes - drug effects; Synaptic Membranes - metabolism; Tetracaine - pharmacology; Torpedo; Torpedo marmorata; USA, Washington
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi034485q

The selectivity of lipid−protein interaction for spin-labeled phospholipids and gangliosides in nicotinic acetylcholine receptor-rich membranes from Torpedo marmorata has been studied by ESR spectroscopy. The association constants of the spin-labeled lipids (relative to phosphatidylcholine) at pH 8.0 are in the order cardiolipin (5.1) ≈ stearic acid (4.9) ≈ phosphatidylinositol (4.7) > phosphatidylserine (2.7) > phosphatidylglycerol (1.7) > GD1b ≈ GM1 ≈ GM2 ≈ GM3 ≈ phosphatidylcholine (1.0) > phosphatidylethanolamine (0.5). No selectivity for mono- or disialogangliosides is found over that for phosphatidylcholine. Aminated local anesthetics were found to compete with spin-labeled phosphatidylinositol, but to a much lesser extent with spin-labeled stearic acid, for sites on the intramembranous surface of the protein. The relative association constant of phosphatidylinositol was reduced in the presence of the different local anesthetics to the following extents:  tetracaine (55%) > procaine (35%) ≈ benzocaine (30%). For stearic acid, only tetracaine gave an appreciable reduction (30%) in association constant. These displacements represent an intrinsic difference in affinity of the local anesthetics for the lipid−protein interface because the membrane partition coefficients are in the order benzocaine ≫ tetracaine ≈ procaine.