Effects of surfactant apolipoproteins on liposome structure: implications for tubular myelin formation.
Tubular myelin is one of several forms of lung surfactant and may play an important role in its surface activity. To determine possible mechanisms of tubular myelin formation, we studied the effects of purified surfactant proteins (SP-A, SP-B, and SP-C) on large unilamellar dipalmitoylphosphatidylcholine-egg phosphatidylglycerol (7/3; wt/wt) liposomes. We studied different types of membrane interaction induced by the apolipoproteins and correlated these with the observed changes in ultrastructure. Aggregation was assessed by measurement of light absorbance, lysis, and fusion by measurement of the fluorescence emitted by water-soluble and lipid-soluble probes, respectively. Mixtures of the apolipoproteins and liposomes were examined in ultrastructural studies by negative staining and by thin sectioning. We found that each protein had a pronounced and distinct effect on liposome structure. SP-A caused aggregation, whereas SP-B and SP-C also caused extensive leakage of liposome contents (lysis) and some degree of lipid mixing (fusion). The disruptive effects of SP-B and to a lesser extent those of SP-C were correlated by negative staining with the appearance of bilayer disks, which tended to aggregate into large sheets. There was a marked synergy between SP-A and SP-B in the process of membrane fusion in the presence of calcium, which correlated with an early (10 min) and extensive rearrangement of the structures seen by electron microscopy followed by a delayed (24 h) appearance of small amounts of tubular myelin.
Poulain
F R
FR
Department of Anatomy and Pediatrics, University of California, San Francisco 94143.
Allen
L
L
Williams
M C
MC
Hamilton
R L
RL
Hawgood
S
S
D000818:Animals; D001053:Apolipoproteins / Q000737:chemistry* / Q000648:ultrastructure*; D001059:Apoproteins / Q000737:chemistry* / Q000648:ultrastructure*; D001688:Biological Products; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008081:Liposomes; D008854:Microscopy, Electron / Q000379:methods; D009186:Myelin Sheath; D010713:Phosphatidylcholines / Q000737:chemistry*; D010715:Phosphatidylglycerols / Q000737:chemistry*; D011506:Proteins; D011510:Proteolipids / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure*
D001053:Apolipoproteins; D001059:Apoproteins; D001688:Biological Products; D008081:Liposomes; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D011506:Proteins; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; C046616:pulmonary surfactant apoprotein; C018167:tubular myelin