Effects of platelet activating factor on calcium-lipid interactions and lateral phase separations in phospholipid...

Recent studies localizing the inflammatory mediator, platelet activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine), to the membranes of stimulated neutrophils, raise the possibility that PAF may, in addition to its activities as a mediator, alter the physical properties of membranes. This, and the increasing evidence that calcium-lipid interactions may have central importance in membrane organizational structure and in functions of cell homeostasis and stimulus-response coupling, prompted us to study the effects of PAF on calcium-lipid interactions in lipid vesicles. Using fluorescence polarization of dansylated probes located in the glycerol portion of the membrane bilayer, PAF (at a concentration as low as 1 mol%) was shown to reduce membrane rigidification significantly during calcium-induced lateral phase separations. This effect of PAF was structurally dependent on both the 1-position alkyl linkage and the 2-position acetyl group as shown by studies of related lipid analogs. Furthermore, using a self-quenching probe, it was shown that inhibition of lateral phase separation did not account for this reduction in the calcium-induced membrane rigidification attributed to PAF. Data suggest that PAF at low concentrations may alter phospholipid head packing and, thereby, change membrane surface features during calcium-lipid interactions, effects which may ultimately explain some of its biological actions.