Reconstitution of tubular myelin from synthetic lipids and proteins associated with pig pulmonary surfactant.
To analyze the mechanism of formation of tubular myelin (TM), we reconstituted TM from synthetic lipids and two surfactant-associated proteins (SP15 and SP35). SP15 was extracted from lyophilized pig pulmonary surfactant with 5% Triton X-100 and purified by DEAE-cellulose, CM-cellulose, and affinity chromatography with a specific antibody. SP35 was extracted from the precipitate of the 5% Triton X-100 extraction with pH 10 borate buffer and purified by DEAE-cellulose column chromatography. Lipid-SP15 complex was formed by a detergent dialysis method using octylglucopyranoside, and to this complex were added various concentrations of SP35 at 37 degrees C. Structures similar to TM were formed when lipid-SP15 complex containing dipalmitoylphosphatidylcholine:phosphatidylglycerol from egg lecithin (2:1) and SP15 (lipid/protein, 5:1) was incubated with SP35 at concentrations of 0.15 to 0.22 mg/ml in CaCl2-containing buffer. At higher concentrations of SP35, many six-sided lattices were formed; the addition of EDTA abolished the formation of these lattice structures. The results suggest that SP15 and SP35 have an important function in the structural organization of lipid membranes to form lattices.
Suzuki
Y
Y
Department of Molecular Pathology, Kyoto University, Japan.
Fujita
Y
Y
Kogishi
K
K
D000818:Animals; D001688:Biological Products; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000378:metabolism*; D066298:In Vitro Techniques; D050356:Lipid Metabolism; D008168:Lung / Q000378:metabolism*; D008854:Microscopy, Electron; D009186:Myelin Sheath; D011506:Proteins; D011510:Proteolipids / Q000378:metabolism*; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000378:metabolism*; D013552:Swine
D001688:Biological Products; D006023:Glycoproteins; D011506:Proteins; D011510:Proteolipids; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; C018167:tubular myelin