Lanthanide(III)-phosphatidic acid complexes: binding site heterogeneity and phase separation

Abstract

The luminescent lanthanides are potentially useful probes of cation-induced events involving phospholipid membranes. In this work, the spectroscopic properties of Tb3+, Ce3+ and Eu3+ are shown to be complementary in defining three forms of complex with phosphatidic acid vesicles. Ce3+, in particular, is useful for studying dilute cation-lipid complexes because it has strong excitation bands in the near ultraviolet. In addition to providing a means for detecting chemically distinct forms of lanthanide-lipid complexes, the luminescence can be used to monitor cation-induced lateral segregation. Ce3+ to Tb3+ energy transfer was observed at lanthanide levels as low as 1:1000 Ln3+/phosphatidic acid, indicating clustering or phase separation. Initial clustering occurs on a subsecond timescale, followed by a much slower aggregation continuing for several minutes to hours. Addition of a chelator results in slow release of the lanthanides. In the case of the dioleoylphosphatidic acid complexes, release is bimodal and indicative of cation entrapment; dimyristoylphosphatidic acid complexes exhibit this behavior only at high temperatures. These observations are consistent with the relative tendencies of these two lipids to form the HII phase. This work sets the foundation for experiments designed to determine the size of nucleation sites for cation-induced events such as intramembrane inverted micelle formation and membrane fusion.