concanavalin-a and 1-2-dipalmitoylphosphatidylglycerol

The interaction of the pulmonary surfactant protein SP-A fluorescently labeled with Texas Red (TR-SP-A) with monolayers of dipalmitoylphosphatidylcholine (DPPC) and DPPC/dipalmitoylphosphatidylglycerol 7:3 w/w has been investigated. The monolayers were spread on aqueous subphases containing TR-SP-A. TR-SP-A interacted with the monolayers of DPPC to accumulate at the boundary regions between liquid condensed (LC) and liquid expanded (LE) phases. Some TR-SP-A appeared in the LE phase but not in the LC phase. At intermediate surface pressures (10-20 mN/m), the protein caused the occurrence of more, smaller condensed domains, and it appeared to be excluded from the monolayers at surface pressure in the range of 30-40 mN/m. TR-SP-A interaction with DPPC/dipalmitoylphosphatidylglycerol monolayers was different. The protein did not appear in either LE or LC but only in large aggregates at the LC-LE boundary regions, a distribution visually similar to that of fluorescently labeled concanavalin A adsorbed onto monolayers of DPPC. The observations are consistent with a selectivity of interaction of SP-A with DPPC and for its accumulation in boundaries between LC and LE phase.

Topics: 1,2-Dipalmitoylphosphatidylcholine; 4-Chloro-7-nitrobenzofurazan; Affinity Labels; Animals; Bronchoalveolar Lavage Fluid; Concanavalin A; Kinetics; Liposomes; Phosphatidylcholines; Phosphatidylglycerols; Pressure; Proteolipids; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants; Spectrometry, Fluorescence; Surface Properties; Swine; Xanthenes

A parallel plate flow chamber with defined wall shear rates was developed in order to study and simulate cellular adhesion to biological membranes as mediated by lectin/carbohydrate interactions. Planar bilayers containing clustered areas of various long-chain alkyl mannosides as carbohydrate ligands and supported on transparent materials were used as model membranes. Their interaction with liposomes bearing Concanavalin A as model cells was observed fluorimetrically by confocal laser scanning microscopy. The use of supported membranes made it possible to study the dependence of adhesion upon different physicochemical parameters of membranes. The liposomes of this model were able to simulate the lectin-mediated adhesion of cells in a shear flow. Once specific receptor-mediated adhesion had taken place, liposomes tended to attach irreversibly to the membrane. This could be avoided by employing lipid compositions which represent a special balance between charged and polyethylene glycol-coupled lipids. This is discussed in term of the interplay between the various attractive and repulsive forces at membrane surfaces. The dependence of liposome adhesion upon the shear rate could be detected. These results were used to evaluate binding forces between lectin-bearing liposomes and ligand-containing planar bilayers.

Topics: Cell Adhesion; Cell Membrane; Concanavalin A; Liposomes; Mannosides; Methylmannosides; Microscopy, Confocal; Phosphatidylglycerols; Polyethylene Glycols; Rheology