Lamellar bodies of cultured human fetal lung: content of surfactant protein A (SP-A), surface film formation and structural transformation in vitro.
Lamellar bodies were isolated from dexamethasone and T3-treated explant cultures of human fetal lung, using sucrose density-gradient centrifugation. We examined their content of surfactant apoprotein A (SP-A), and their ability to form surface films and to undergo structural transformation in vitro. SP-A measured by ELISA composed less than 2% of total protein within lamellar bodies; this represented, as a minimum estimate, a 2-12-fold enrichment over homogenate. One- and two-dimensional gel electrophoresis also suggested that SP-A was a minor protein component of lamellar bodies. Adsorption of lamellar bodies to an air/water interface was moderately rapid, but accelerated dramatically upon addition of exogenous SP-A in ratios of 1:2-16 (SP-A:phospholipid, w/w). Similar adsorption patterns were seen for lamellar bodies from fresh adult rat and rabbit lung. Lamellar bodies incubated under conditions that promote formation of tubular myelin underwent structural rearrangement only in the presence of exogenous SP-A, with extensive formation of multilamellate whorls of lipid bilayers (but no classical tubular myelin lattices). We conclude that lamellar bodies are enriched in SP-A, but have insufficient content of SP-A for structural transformation to tubular myelin and rapid surface film formation in vitro.
Froh
D
D
Department of Pediatrics, University of California, San Francisco.
Ballard
P L
PL
Williams
M C
MC
Gonzales
J
J
Goerke
J
J
Odom
M W
MW
Gonzales
L W
LW
D002458:Cell Fractionation; D002478:Cells, Cultured; D003600:Cytosol / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005333:Fetus; D006801:Humans; D008168:Lung / Q000378:metabolism / Q000648:ultrastructure*; D008715:Methionine / Q000378:metabolism; D008854:Microscopy, Electron; D015388:Organelles / Q000648:ultrastructure*; D010743:Phospholipids / Q000032:analysis; D011510:Proteolipids / Q000032:analysis* / Q000096:biosynthesis; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000032:analysis* / Q000096:biosynthesis; D013347:Subcellular Fractions / Q000032:analysis; D013462:Sulfur Radioisotopes
D010743:Phospholipids; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D013462:Sulfur Radioisotopes; D008715:Methionine