1-palmitoyl-2-oleoylphosphatidylcholine and 1-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine

The enhanced permeability of lipid bilayer membranes at their gel-to-liquid phase transition has been explained using a "bilayer lipid heterogeneity" model, postulating leaky interfacial regions between still solid and melting liquid phases. The addition of lysolipid to dipalmitoylphosphatidylcholine bilayers dramatically enhances the amount of, and speed at which, encapsulated markers or drugs are released at this, already leaky, phase transition through these interfacial regions. To characterize and attempt to determine the mechanism behind lysolipid-generated permeability enhancement, dithionite permeability and doxorubicin release were measured for lysolipid and non-lysolipid, containing membranes. Rapid release of contents from lysolipid-containing membranes appears to occur through lysolipid-stabilized pores rather than a simple enhancement due to increased drug solubility in the bilayer. A dramatic enhancement in the permeability rate constant begins about two degrees below the calorimetric peak of the thermal transition, and extends several degrees past it. The maximum permeability rate constant coincides exactly with this calorimetric peak. Although some lysolipid desorption from liquid state membranes cannot be dismissed, dialyzation above T(m) and mass spectrometry analysis indicate lysolipid must, and can, remain in the membrane for the permeability enhancement, presumably as lysolipid stabilized pores in the grain boundary regions of the partially melted solid phase.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Biophysics; Calorimetry; Calorimetry, Differential Scanning; Dithionite; Doxorubicin; Ions; Lipid Bilayers; Lipids; Liposomes; Mass Spectrometry; Membranes, Artificial; Models, Chemical; Permeability; Phosphatidylcholines; Spectrophotometry; Temperature; Time Factors

This paper reports the detailed composition of molecular species of the phosphatidylcholines (PCs) in pulmonary surfactant from calves. PC isolated by thin-layer chromatography (TLC) was converted to benzoylated diradyl glyceride derivatives, which were separated by TLC according to linkage group. Quantification of linkage groups by analysis of total fatty acid content demonstrated that surfactant PC contained 97.2% diacyl, 2.4% alkyl-acyl, and 0.4% alkenyl-acyl compounds. The diacyl and alkyl-acyl diglyceride derivatives were separated into individual molecular species by high-performance liquid chromatography. Four major species constituted 87% of the diacyl compounds. Dipalmitoyl phosphatidylcholine (DPPC) was the most abundant constituent, contributing 41% of the total PC. A second disaturated species, palmitoyl-myristoyl phosphatidylcholine (PMPC), also contributed an additional 12% of total PC. At least 65% of PMPC occurred as the 1-palmitoyl-2-myristoyl/isomer, which has a lower melting point than the 1-myristoyl-2-palmitoyl compound. These results show that most of pulmonary surfactant PC is a relatively simple mixture, that numerous minor compounds are present in small but possibly important amounts, and that in surfactant from calves, the widely reported estimate that DPPC constitutes 60% of surfactant PC is too large by 50%.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Animals; Cattle; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Fatty Acids; Hydrolysis; Phosphatidylcholines; Phospholipases A; Pulmonary Surfactants

Lipase-catalyzed oxygen exchange between 13,16-cis,cis-docosadienoic acid and water in liquid-expanded monolayers with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine exhibits abrupt, lipid composition-dependent changes in extent and mechanism [e.g., Muderhwa, J. M. and Brockman, H. L. (1992) J. Biol. Chem. 267, 24184-24192]. The critical nature of this transition suggests possible lateral phase separation of the lipids. This has been addressed by substituting for either lipid species one which can exist in more condensed monolayer states. Analysis of phase transition surface pressures as a function of lipid composition shows that each set of fatty acid-phosphatidylcholine mixtures exhibits a finite range of miscibility in liquid-expanded monolayers. These results strongly suggest that 13,16-cis,cis-docosadienoic acid and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine are miscible under the conditions of the oxygen-exchange experiments. Furthermore, to address more directly the relation of lateral lipid phase separation to lipase regulation, oxygen exchange catalyzed by pancreatic carboxylester and triglyceride lipases was studied using mixed monolayers of [18O]2-docosadienoic acid and 1-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine. These lipids are miscible in the liquid-expanded state at all compositions. The lipid composition dependencies of both the extent and mechanism of lipase-catalyzed oxygen exchange were essentially identical to those obtained earlier. Thus, lateral lipid phase separation is not required for the critical transition in substrate accessibility to lipases. This finding supports a percolation-based model of lipase regulation within a single surface phase and suggests the "topo-temporal" regulation of lipid-mediated signaling in cells.

Topics: Chemical Phenomena; Chemistry, Physical; Fatty Acids; Fatty Acids, Unsaturated; Lipase; Lysophosphatidylcholines; Oxygen; Phosphatidylcholines