Physical properties and surface activity of surfactant-like membranes containing the cationic and hydrophobic peptide KL4

Surfactant-like membranes containing the 21-residue peptide KLLLL-KLLLLKLLLLKLLLLK (KL4), have been clinically tested as a therapeutic respiratory distress syndrome in premature infants. The aims of this study were to investigate the interactions between the KL4 peptide and lipid bilayers, and the role of both the lipid composition and KL4 structure on the surface adsorption activity of KL4-containing membranes. We used bilayers of three-component systems [1,2-dipalmitoyl-phosphatidylcholine/1-palmitoyl-2-oleoyl-phosphatidylglyceroVpalmitic acid (DPPC/ POPG/PA) and DPPC/1-palmitoyl-2-oleoyl-phosphatidylcholine (POPQ/ PA] and'binary lipid mixtures of DPPC/POPG and DPPC/PA to examine the specific interaction of KL4 with POPG and PA. We found that, at low peptide concentrations, KL4 adopted a predominantly alpha-helical secondary structure in POPG- or POPC-containing membranes, and a beta-sheet structure in DPPC/PA vesicles. As the concentration of the peptide increased, KL4 interconverted to a beta-shect structure in DPPC/POPG/PA or DPPC/POPC/PA vesicles. Ca2+ favored alpha <-> beta interconversion. This conformational flexibility of KL4 did not influence the surface adsorption activity of KL4-containing vesicles. KL4 showed a concentration-dependent ordering effect on POPG- and POPC-containing membranes, which could be linked to its surface activity. In addition, we found that the physical state of the membrane had a critical role in the surface adsorption process. Our results indicate that the most rapid surface adsorption takes place with vesicles showing well-defined solid/fluid phase co-existence at temperatures below their gel to fluid phase transition temperature, such as those of DPPC/POPG/PA and DPPC/POPC/PA. In contrast, more fluid (DPPC/POPG) or excessively rigid (DPPC/PA) KL4-containing membranes fail in their ability to adsorb rapidly onto and spread at the air water interface.