A method to detect leakage of DNA intercalators through liposome membranes.
A method to detect leakage of water-soluble intercalators such as ethidium bromide and propidium iodide from liposomes (egg-lecithin and egg-lecithin/cholesterol) by means of fluorescence is presented. The addition of excess DNA outside the liposomes brings about intercalation of released dye with the DNA. The increase in fluorescence intensity that results from the dye-DNA complex is used to detect the degree of dye release. Titration and surfactant-induced leakage measurements display the suitability of the method. Spontaneous leakage measurements show that propidium has much lower leaking rates than those of ethidium. This is probably due to the less lipophilicity of propidium as estimated from the partition coefficients between octanol and medium. Stabilizing the egg-lecithin membranes with cholesterol strongly reduces the leaking rates for ethidium while no effect is detected for the slow leaking rates of propidium. Lowering the temperature from 25 to 6 degrees C results in lower leaking rates for both dyes.
Silvander
M
M
Department of Physical Chemistry, Uppsala University, Sweden.
Edwards
K
K
D002784:Cholesterol; D004247:DNA / Q000032:analysis*; D004996:Ethidium / Q000032:analysis* / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis; D007364:Intercalating Agents / Q000032:analysis* / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D010713:Phosphatidylcholines; D011419:Propidium / Q000032:analysis* / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D007364:Intercalating Agents; D008081:Liposomes; D010713:Phosphatidylcholines; D011419:Propidium; D004247:DNA; D002784:Cholesterol; D004996:Ethidium