Kinetics of phospholipid insertion into monolayers containing the lung surfactant proteins SP-B or SP-C.
The lung surfactant proteins SP-B and SP-C are pivotal for fast and reversible lipid insertion at the air/liquid interface, a prerequisite for functional lung activity. We used a model system consisting of a preformed monolayer at the air/liquid interface supplemented with surfactant protein SP-B or SP-C and unilamellar vesicles injected into the subphase of a film balance. The content of SP-B or SP-C was similar to that found in lung lavage. In order to elucidate distinct steps of lipid insertion, we measured the time-dependent pressure increase as a function of the initial surface pressure, the temperature and the phosphatidylglycerol content by means of surface tension measurements and scanning force microscopy (SFM). The results of the film balance study are indicative of a two-step mechanism in which initial adsorption of vesicles to the protein-containing monolayer is followed by rupture and integration of lipid material. Furthermore, we found that vesicle adsorption on a preformed monolayer supplemented with SP-B or SP-C is strongly enhanced by negatively charged lipids as provided by DPPG and the presence of Ca2+ ions in the subphase. Hence, long-range electrostatic interactions are thought to play an important role in attracting vesicles to the surface, being the initial step in replenishment of lipid material. While insertion into the monolayer is independent of the type of protein SP-B or SP-C, initial adsorption is faster in the presence of SP-B than SP-C. We propose that the preferential interaction between SP-B and negatively charged DPPG leads to accumulation of negative charges in particular regions, causing strong adhesion between DPPG-containing vesicles and the monolayer mediated by Ca2+ ions, which eventually causes flattening and rupture of attached liposomes as observed by in situ SFM.
Ross
Michaela
M
Institut für Biochemie, Westfälische Wilhelms-Universität, Münster, Germany.
Krol
Silke
S
Janshoff
Andreas
A
Galla
Hans-Joachim
HJ
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008081:Liposomes; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D010715:Phosphatidylglycerols / Q000235:genetics / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*; D037701:Pulmonary Surfactant-Associated Protein B / Q000737:chemistry / Q000378:metabolism*; D037721:Pulmonary Surfactant-Associated Protein C / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D055672:Static Electricity; D013552:Swine; D013696:Temperature
D008081:Liposomes; D010715:Phosphatidylglycerols; D010743:Phospholipids; D037701:Pulmonary Surfactant-Associated Protein B; D037721:Pulmonary Surfactant-Associated Protein C; D011994:Recombinant Proteins; D015060:1,2-Dipalmitoylphosphatidylcholine; C030345:1,2-dipalmitoylphosphatidylglycerol