Suppression of Lα/Lβ Phase Coexistence in the Lipids of Pulmonary Surfactant

To determine how different constituents of pulmonary surfactant affect its phase behavior, we measured wideangle x-ray scattering (WAXS) from oriented bilayers. Samples contained the nonpolar and phospholipids (N&PL) obtained from calf lung surfactant extract (CLSE), which also contains the hydrophobic surfactant proteins SP-B and SP-C. Mixtures with different ratios of N&PL and CLSE provided the same set of lipids with different amounts of the proteins. At 37 degrees C, N&PL by itself forms coexisting L-alpha, and L-beta phases. In the L-beta structure, the acyl chains of the phospholipids occupy an ordered array that has melted by 40 degrees C. This behavior suggests that the L-beta, composition is dominated by dipalmitoyl phosphatidylcholine (DPPC), which is the most prevalent component of CLSE. The L-beta chains, however, lack the tilt of the L-beta, phase formed by pure DPPC. At 40 degrees C, WAXS also detects an additional diffracted intensity, the location of which suggests a correlation among the phospholipid headgroups. The mixed samples of N&PL with CLSE show that increasing amounts of the proteins disrupt both the L-beta phase and the headgroup correlation. With physiological levels of the proteins in CLSE, both types of order are absent. These results with bilayers at physiological temperatures indicate that the hydrophobic surfactant proteins disrupt the ordered structures that have long been considered essential for the ability of pulmonary surfactant to sustain low surface tensions. They agree with prior fluorescence micrographic results from monomolecular films of CLSE, suggesting that at physiological temperatures, any ordered phase is likely to be absent or occupy a minimal interfacial area.

Automated summary

This study used wide-angle x-ray scattering from oriented bilayers to explore the effects of different constituents of pulmonary surfactant on its phase behavior. It was found that hydrophobic surfactant proteins disrupt the ordered structures of pulmonary surfactant, which are crucial for maintaining low surface tensions.