We have shown previously that lateral compression of pulmonary surfactant monolayers initially induces separation of two phases but that these remix when the films become more dense (1). In the studies reported here, we used fluorescence microscopy to examine the role of the different surfactant constituents in the remixing of the separated phases. Subfractions containing only the purified phospholipids (PPL), the surfactant proteins and phospholipids (SP&PL), and the neutral and phospholipids (N&PL) were obtained by chromatographic separation of the components in extracted calf surfactant (calf lung surfactant extract, CLSE). Compression of the different monolayers produced nonfluorescent domains that emerged for temperatures between 20 and 41°C at similar surface pressures 6-8 mN/m higher than values observed for dipalmitoyl phosphatidylcholine (DPPC), the most prevalent component of pulmonary surfactant. Comparison of the different preparations showed that the neutral lipid increased the total nonfluorescent area at surface pressures up to 25 mN/m but dispersed that total area among a larger number of smaller domains. The surfactant proteins also produced smaller domains, but they had the opposite effect of decreasing the total nonfluorescent area. Only the neutral lipids caused remixing. In images from static monolayers, the domains for N&PL dropped from a maximum of 26 ± 3% of the interface at 25 mN/m to 4 ± 2% at 30 mN/m, similar to the previously reported behavior for CLSE. During continuous compression through a narrow range of pressure and molecular area, in N&PL, CLSE, and mixtures of PPL with 10% cholesterol, domains became highly distorted immediately prior to remixing. The characteristic transition in shape and abrupt termination of phase coexistence indicate that the remixing caused by the neutral lipids occurs at or close to a critical point. † © 1999 American Chemical Society * To whom correspondence should be addressed at Mail Code OHSU, Portland,..
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Author ManuscriptBiochemistry. Author manuscript; available in PMC 2012 December 08. The phase behavior of biological phospholipid films and membranes may be important in at least two respects. First, the different physical characteristics of the different phases can alter processes such as lateral diffusion. Second, phase separation can maintain components in distinct compartments and provide a means of organizing biological processes (2). Both of these issues may be important for the function of pulmonary surfactant. This complex mix of lipids and proteins coats the thin liquid layer that lines the lung's alveolar air spaces. When compressed by decreasing alveolar surface areas during normal exhalation, the surfactant film achieves very high densities, well above equilibrium values. Measurements in situ show that the surfactant films lower the surface tension of the air-liquid interface to very low levels (3) and in doing so stabilize the alveoli. Only films with the physical characteristics of a highly ordered phase se...