Calcium-induced lipid phase separations and interactions of phosphatidylcholine/anionic phospholipid vesicles. Fluorescence studies using carbazole-labeled and brominated phospholipids.
A novel method that uses a carbazole-labeled fluorescent phosphatidylcholine, which partitions preferentially into liquid-crystalline lipid domains, to monitor the kinetics and the extents of thermotropic and ionotropic lateral phase separations in vesicles combining brominated and nonbrominated phosphatidylcholines (PCs), phosphatidic acids (PAs), and phosphatidylserines (PSs) is described. The calcium-induced segregation of several nonbrominated PA species in liquid-crystalline brominated PC bilayers behaves as a well-defined lateral phase separation; the residual solubility of the PA component in the PC-rich phase in the presence of calcium can vary severalfold depending on the PA acyl chain composition. PC/PS mixtures show a pronounced tendency to form metastable solutions in the presence of calcium, particularly when they contain less than equimolar proportions of PS. This metastability is not readily relaxed by repeated freeze-thawing of vesicles in the presence of calcium, by avidin-mediated contacts between PC/PS vesicles containing biotinylated lipids, or by calcium-induced lateral segregation of PA in the same vesicles. Different PS species exhibit different apparent residual solubilities in liquid-crystalline PC bilayers, ranging from less than 10 mol % for dimyristoyl-PS to ca. 45 mol% for dioleoyl-PS, after prolonged incubations of PC/PS multilamellar vesicles with excess calcium. Results are presented, obtained by using the above lipid-segregation assay and parallel assays of intervesicle lipid mixing, that raise questions concerning the relevance of the equilibrium behavior of calcium-treated PS/PC mixtures to the relatively rapid interactions (fusion and lipid mixing) of PC/PS vesicles that follow initial exposure to calcium.
Silvius
J R
JR
Department of Biochemistry, McGill University, Montréal, Québec, Canada.
D001966:Bromine; D002118:Calcium; D002227:Carbazoles; D007700:Kinetics; D008081:Liposomes; D008433:Mathematics; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D012997:Solvents; D013050:Spectrometry, Fluorescence / Q000379:methods; D013329:Structure-Activity Relationship
D002227:Carbazoles; D008081:Liposomes; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D012997:Solvents; D001966:Bromine; D002118:Calcium