Interaction of the lipid and protein components of pulmonary surfactant Role of phosphatidylglycerol and calcium

We investigated the interaction of a major apolipoprotein of pulmonary surfactant with mixtures of lipids analogous to those found in natural surfactant. The apolipoprotein was extracted from canine surfactant and was purified to about 90% homogeneity. The apolipoprotein was mixed with liposomes of lipids in buffers containing 0.1 M sodium chloride and 3 mM calcium chloride at 22°C for 2 h or 37°C for 30 min. Two fractions were separated by centrifugation in sucrose density gradients at 15 000 rev./min. One was comprised of an aggregated, relatively high density recombinant lipoprotein which sedimented to a position toward the bottom of the centrifuge tube; the other remained at the top of the centrifuge tube and was mainly comprised of unbound lipid. The amount of lipid recovered as a sedimenting lipoprotein was dependent upon its composition. Those mixtures of lipids which contained dipalmitoyl phosphatidylglycerol formed sedimenting complexes which comprised 14% to 53% of the recovered lipid; those without phosphatidylglycerol formed such aggregates with less than 13% of the available lipid. Moreover, the lipid-to-protein stoichiometry of the recombinant was also dependent upon phosphatidylglycerol, and lipids containing this phospholipid displayed enhanced binding at a critical concentration of lipid which varied with temperature and composition. Calcium was required to form the sedimenting complex at 37°C. These results suggest that phosphatidylglycerol may be involved in the formation of a micelle-like complex, the stoichiometry of which is regulated over a narrow range of lipid concentration, and the structure of which involves calcium. The physiological advantage of forming this complex has not been determined. We found, however, that lipids containing phosphatidylglycerol absorbed more rapidly to an air/liquid interface than did those without. This rate of adsorption was further increased after interaction with the apolipoprotein.