Pulmonary Surfactant Protein SP-A with Phospholipids in Spread Monolayers at the Air-Water Interface

Abstract

Spread monolayers of pulmonary surfactant protein SP-A, alone or mixed with phospholipid(s), were formed at the air-water interface. Binary monolayers of SP-A plus dipalmitoylphosphatidylcholine (DPPC) showed positive deviations from ideal behavior of the mean areas in the films consistent with partial miscibility and interaction between the protein and lipid. During compression of SP-A/DPPC films which contained > or = 5 wt % SP-A, properties were displayed which were consistent with the protein being partially squeezed out at surface pressures of about 30 mN/m. Some protein appeared to remain in the monolayers even when they were compressed to high surface pressures of about 65-70 mN/m, and it was possibly included in the collapse phase(s) that was (were) formed at 72 mN/m. During dynamic cyclic compression-expansion of SP-A/DPPC monolayers initially formed at low surface pressures, SP-A enhanced the respreading of the films compressed beyond collapse compared to the respreading after collapse of films containing DPPC alone. Spread monolayers of SP-A plus either dipalmitoylphosphatidylglycerol (DPPG) or a mixture of DPPC plus DPPG (7:3, mol/mol) displayed additivity of the mean areas in the films, consistent with complete immiscibility (or ideal miscibility, an unlikely effect) between the protein and lipid components. Electrostatic repulsion between SP-A and DPPG, both negatively charged at physiological pH, possibly governed the behavior of these lipid-protein films. Calcium ions in the subphase did not alter the properties of SP-A/DPPC films, whereas they improved the ability of SP-A to mix with DPPG and DPPC/DPPG. Binding of calcium to the negatively charged DPPG and SP-A may account for association of the protein with DPPG and DPPC/DPPG in the monolayers in the presence of the divalent ions.