Reassembly of lipid-protein complexes of pulmonary surfactant. Proposed mechanism of interaction.
We studied the interaction at 37 degrees C between a major apolipoprotein of pulmonary surfactant and 11 mixtures of lipids. The experiments were carried out in the presence of either 3 mM Ca2+ or 10 mM EDTA. The amount of apolipoprotein associated with lipid was independent of Ca2+. However the binding was sensitive to the percentage of gel-state lipid in the vesicles, and the amount of apolipoprotein in the recombinant lipoprotein complex decreased as the percentage of fully saturated phospholipid was reduced. Maximum association of the apolipoprotein occurred with lipid vesicles containing 85% 1,2-dipalmitoyl-sn-glycero-3-phosphocholine and 15% 1,2-dipalmitoyl-sn-glycero-3-phospho-1-glycerol or 1,2-dipalmitoyl-sn-glycerol. Fluorescence measurements on the apolipoprotein indicated that the tryptophan side chains were in a relatively hydrophobic environment, and that the wavelength of maximum fluorescence emission was not changed upon the binding of lipid. The results suggest that the principal mode of interaction between the apolipoprotein and lipids of surfactant is hydrophobic bonding. The most extensive binding occurs with lamellar lipids in a gel that would be expected to have inhomogeneities in packing density due to the presence of acidic phospholipids or other glycerolipids. The role of Ca2+ in this interaction has not been fully determined. Although it is not needed to effect the binding of the lipids and the apolipoprotein, it does influence the physical state of the complex, and possibly the stoichiometry of lipid to protein. Some of the processes mediated by Ca2+ in this interaction may be analogous to those observed in membrane fusion. Thus, Ca2+ probably causes segregation of the lamellar phospholipids into domains, inducing vesicular disruption and fusion. This lipid aggregates about hydrophobic sites on the protein, thereby forming high molecular weight reassembly complexes.
D000818:Animals; D001053:Apolipoproteins / Q000378:metabolism*; D001054:Apolipoproteins A; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008970:Molecular Weight; D010715:Phosphatidylglycerols / Q000378:metabolism*; D011487:Protein Conformation; D011663:Pulmonary Surfactants / Q000378:metabolism*
D001053:Apolipoproteins; D001054:Apolipoproteins A; D046911:Macromolecular Substances; D010715:Phosphatidylglycerols; D011663:Pulmonary Surfactants; D002118:Calcium; C030345:1,2-dipalmitoylphosphatidylglycerol