Interaction of local anesthetics with small phospholipid vesicles investigated by proton NMR spectroscopy
Tóm tắt
The effects of the local anesthetics tetracaine, procaine (both charged at pH 6), and benzocaine (uncharged) on phospholipid liposomes have been investigated by 500 MHz 1H NMR Spectroscopy. All the drugs reverse the Pr3+ induced shifts of phospholipid resonances in the same sequence as they are shifted by addition of Pr3+: choline POCH2- > choline-CH2N > choline-N(CH3)3 > glycerol Β > glycerol γ > acyl C2 > acyl C3. The drug effects result from incorporation of positive charges (tetracaine and procaine) and from the induction of a conformational change of the phospholipid head group via an action on the lipid glycerol backbone (benzocaine). From titration experiments with tetracaine on liposomes containing Pr3+ inside and outside is derived that the drug passes the bilayer by transverse diffusion. Tetracaine partitions outside/inside at a ratio of 2∶1. Changes in linewidths of the drug resonances when incorporated into the liposomes allow the conclusion that the tetracaine molecule is located in an elongated way between the lipid acyl chains with its nitrogen group near the glycerol backbone. Benzocaine, showing strong effects on the line shapes of the protons on C2 and C3 of the lipid acyl chains is also located near the glycerol backbone, the region with the strongest hydrophobic forces.
Tài liệu tham khảo
Boulanger Y, Schreier S, Smith ICP (1981) Molecular details of anesthetic-lipid interaction as seen by deuterium and phosphorus-31 nuclear magnetic resonance. Biochemistry 20: 6824–6830
Browning JL, Akutsu H (1982) Local anesthetics and divalent cations have the same effect on the headgroups of phosphatidylcholine and phosphatidylethanolamine. Biochim Biophys Acta 684: 172–178
Bystrov VF, Dubrovina NI, Barsukov LR, Bergelson LD (1971) NMR differentiation of the internal and external phospholipid membrane surfaces using paramagnetic Mn2+ and Eu3+ ions. Chem Phys Lipids 6: 343–350
Dittmer JC, Lester RL (1964) A simple, specific spray for the detection of phospholipids on thin-layer chromatograms. J Lipid Res 37: 126–127
Eriksson LEG, Westman J (1981) Interaction of some charged amphiphilic drugs with phosphatidylcholine vesicles. A spin label study of surface potential effects. Biophys Chem 13: 253–264
Fernandez MS, Cerbon J (1973) The importance of the hydrophobic interactions of local anesthetics in the displacement of polyvalent cations from artificial lipid membranes. Biochim Biophys Acta 298: 8–14
Gee A, Deitz VR (1953) Determination of phosphate by differential spectrophotometry. Anal Chem 25: 1320–1324
Grasdalen H, Eriksson LEG, Westman J, Ehrenberg A (1977) Surface potential effects on metal ion binding to phosphatidylcholine membranes. 31P NMR study of lanthanide and calcium ion binding to egg-yolk lecithin vesicles. Biochim Biophys Acta 469: 151–162
Hauser H (1981) The polar group conformation of 1,2-dialkyl phosphatidylcholines. An NMR study. Biochim Biophys Acta 646: 203–210
Hauser H, Dawson RMC (1968) The displacement of calcium ions from phospholipid monolayers by pharmacologically active and other organic bases. Biochem J 109: 909–916
Hauser H, Phillips MC, Levine BA, Williams RJP (1975) Ion-binding to phospholipids. Interaction of calcium and lanthanide ions with phosphatidylcholine (lecithin). Eur J Biochem 58: 133–144
Hauser H, Guyer W, Levine B, Skrabal P, Williams RJP (1978) The conformation of the polar group of lysophosphatidylcholine in H2O; conformational changes induced by polyvalent cations. Biochim Biophys Acta 508: 450–463
Haydon DA, Hendry BM, Levinson SR, Requena J (1977) The molecular mechanism of anaesthesia. Nature 268: 356–358
Hille B (1977) Local anesthetics: hydrophilic and hydrophobic pathways for the drug-receptor reaction. J Gen Physiol 69: 497–515
Hunt GRA, Tipping LRH (1980) Trans-bilayer pseudocontact shifts produced by lanthanide ions in the 1H and 31P-NMR spectra of phospholipid vesicular membranes and their temperature variation. J Inorganic Biochem 12: 17–36
Khodorov G, Shishkova L, Peganov E, Revenko S (1976) Inhibition of sodium currents in frog Ranvier node treated with local anesthetics. Role of slow inactivation. Biochim Biophys Acta 433: 409–435
Klein RA (1970) The large-scale preparation of unsaturated phosphatidylcholine from egg-yolk. Biochim Biophys Acta 219: 496–499
Lee AG (1978) Effects of charged drugs on the phase transition temperatures of phospholipid bilayers. Biochim Biophys Acta 514: 95–104
McLaughlin S (1975) Local anesthetics and the electrical properties of phospholipid bilayer membranes. In Fink BR (ed) Molecular mechanisms of anesthesia, progress in anesthesiology, vol 1. Raven Press, New York, pp 193–200
Michaelis L (1980) Location and motion of ubiquinone 50 in liposome membranes. Dissertation (PhD), University of Hull
Michaelis L, Schlieper P (1982) 500 MHz 1H NMR of phospholipid liposomes. FEBS Lett 147: 40–43
Papahadjopoulos D (1970) Phospholipid model membranes. III. Antagonistic effects of Ca2+ and local anesthetics on the permeability of phosphatidylserine vesicles. Biochim Biophys Acta 211: 467–477
Papahadjopoulos D, Jacobsen K, Poste G, Shephard G (1975) Effects of local anesthetics on membrane properties. 1. Change in the fluidity of phospholipid bilayers. Biochim Biophys Acta 394: 504–519
Schlieper P, Medda PK, Kaufmann R (1981) Drug-induced zeta potential changes in liposomes studied by laser Doppler spectroscopy. Biochim Biophys Acta 644: 273–283
Skou JC (1954) Local anaesthetics. VI. Relation between blocking potency and penetration of a monomolecular layer of lipids from nerves. Acta Pharmacol Toxicol 10: 281–337
Yeagle PL, Hutton WC, Martin RB (1977) Molecular dynamics of the local anesthetic tetracaine in phospholipid vesicles. Biochim Biophys Acta 465: 173–178