Mode of interaction of hydrophobic amphiphilic alpha-helical peptide/dipalmitoylphosphatidylcholine with phosphatidylglycerol or palmitic acid at the air-water interface.
Surface pressure (pi)-, surface potential (DeltaV)-, dipole moment (mu( perpendicular))-area (A) isotherms and morphological behavior at the air-water interface were obtained for multicomponent monolayers of two different systems for dipalmitoylphosphatidylcholine (DPPC)/egg-phosphatidylglycerol (PG) (= 68:22, by weight)/Hel 13-5 and DPPC/palmitic acid (PA) (= 90:9, by weight)/Hel 13-5 (Hel 13-5 is a newly designed 18-mer amphiphilic alpha-helical peptide with 13 hydrophobic and 5 hydrophilic amino acid residues). The phase behavior of these model systems was investigated on a subsolution of 0.02 M tris(hydroxymethyl)aminomethane (Tris) buffer (pH 8.4) with 0.13 M NaCl at 298.2 K by employing the Wilhelmy method, the ionizing electrode method, and fluorescence microscopy. Especially, the present study focuses on the interfacial effect of the addition of Hel 13-5 on two binary systems, DPPC/egg-PG and DPPC/PA monolayers, as the substitute for pulmonary surfactant proteins, and on the respective roles of PG and PA for the monolayers in the three-component systems. Constant kink points ( approximately 42 mN m(-1)) clearly appear on the pi-A isotherms, independent of the compositions in the ternary systems, which corresponds to the Hel 13-5 collapse pressure similar to that of SP-B and SP-C as functions in multicomponent monolayers. This implies that Hel 13-5 is squeezed out of ternary monolayers above approximately 42 mN m(-1), resulting in two- to three-dimensional phase transformation. Furthermore, Langmuir isotherms clearly show that Hel 13-5 with egg-PG is squeezed out of the DPPC/egg-PG/Hel 13-5 system, whereas only Hel 13-5 is squeezed out of the DPPC/PA/Hel 13-5 system. Cyclic compression and expansion isotherms of these systems were carried out to confirm the spreading and respreading capacities. In addition, the interfacial behavior of the ternary mixtures has been analyzed by the additivity rule. Morphological examinations and comparisons have verified the interactions of Hel 13-5 with the representative miscible mixture (DPPC/PA system) by fluorescence microscopy. Consequently, distinct morphological variations corresponding to the squeeze-out behavior are observed as a fluorescent contrast recovery. Herein, a new mechanism of the refluorescent phenomenon is proposed by varying the surface composition of Hel 13-5.
Nakahara
Hiromichi
H
Division of Biointerfacial Science, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Lee
Sannamu
S
Sugihara
Gohsuke
G
Shibata
Osamu
O
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry*; D000388:Air; D000595:Amino Acid Sequence; D057927:Hydrophobic and Hydrophilic Interactions; D046911:Macromolecular Substances / Q000737:chemistry; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019308:Palmitic Acid / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D010715:Phosphatidylglycerols / Q000737:chemistry*; D017433:Protein Structure, Secondary; D013501:Surface-Active Agents / Q000737:chemistry; D013816:Thermodynamics; D014867:Water
D046911:Macromolecular Substances; D010455:Peptides; D010715:Phosphatidylglycerols; D013501:Surface-Active Agents; D014867:Water; D015060:1,2-Dipalmitoylphosphatidylcholine; D019308:Palmitic Acid