Binding of imipramine to phospholipid bilayers using radioligand binding assay.
Binding of the tricyclic antidepressant imipramine (IMI) to neutral and negatively charged lipid membranes was investigated using a radioligand binding assay combined with centrifugation or filtration. Lipid bilayers were composed of brain phosphatidylcholine (PC) and phosphatidylserine (PS). IMI binding isotherms were measured up to IMI concentration of 0.5 mmol/l. Due to electrostatic attraction, binding between the positively charged IMI and the negatively charged surfaces of PS membranes was augmented compared to binding to neutral PC membranes. After correction for electrostatic effects by means of the Gouy-Chapman theory, the binding isotherms were described both by surface partition coefficients and by binding parameters (association constants and binding capacities). It was confirmed that binding of IMI to model membranes is strongly affected by negatively charged phospholipids and that the binding is heterogeneous; in fact, weak surface adsorption and incorporation of the drug into the hydrophobic core of lipid bilayer can be seen and characterized. These results support the hypothesis suggesting that the lipid part of biological membranes plays a role in the mechanism of antidepressant action.
Department of Psychiatry, 1st Faculty of Medicine, Charles University, Prague, Czech Republic. zfisar@lf1.cuni.cz
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Fisar
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Fuksová
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Velenovská
D000818:Animals; D000929:Antidepressive Agents, Tricyclic / Q000032:analysis / Q000737:chemistry; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002498:Centrifugation; D005374:Filtration; D007099:Imipramine / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers / Q000032:analysis / Q000737:chemistry*; D008081:Liposomes / Q000032:analysis / Q000737:chemistry*; D046911:Macromolecular Substances / Q000032:analysis / Q000737:chemistry; D008567:Membranes, Artificial; D008956:Models, Chemical; D010713:Phosphatidylcholines / Q000032:analysis / Q000737:chemistry*; D010718:Phosphatidylserines / Q000032:analysis / Q000737:chemistry*; D010743:Phospholipids / Q000032:analysis / Q000737:chemistry; D011869:Radioligand Assay / Q000379:methods*
D000929:Antidepressive Agents, Tricyclic; D008051:Lipid Bilayers; D008081:Liposomes; D046911:Macromolecular Substances; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D010743:Phospholipids; D007099:Imipramine