Isolation, characterization and phospholipid composition of lamellar bodies and subcellular fractions from dog lung.
1. Lamellar body fractions from dog lung can be separated by a procedure based on differential centrifugation before ultracentrifugation onto a discontinuous sucrose gradient. This fraction yields about 1% of total protein from the homogenate. 2. The different fractions obtained in the isolation were assayed for the measurement of four subcellular marker enzymes: beta-N-acetylglucosaminidase, acid phosphatase, 5'-nucleotidase and succinate dehydrogenase. 3. Lamellar bodies were not contaminated by mitochondria (0.7 succinate dehydrogenase relative specific activity), whereas high specific hydrolase activities were found (beta-N-acetylglucosaminidase and 5'-nucleotidase were enriched 1.8- and 2.8-fold, respectively). 4. The chemical criterion was established by measuring the specific components of lamellar bodies. The lamellar bodies have the highest phospholipid/protein ratio (0.35); cholesterol/protein ratio (0.15) and the highest phosphatidylglycerol percentages (7.9%). 5. The phospholipid composition of lamellar bodies is distributed among phosphatidylcholine (64.5%), phosphatidylethanolamine (11%), phosphatidylglycerol (7.9%), sphingomyelin (4%), phosphatidylserine and phosphatidylinositol (3%), respectively. The remainder were considered as trace amounts (less than 1%).
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D004285:Dogs; D008168:Lung / Q000032:analysis / Q000648:ultrastructure*; D009940:Organoids / Q000032:analysis / Q000648:ultrastructure*; D010743:Phospholipids / Q000032:analysis*; D013347:Subcellular Fractions / Q000032:analysis / Q000648:ultrastructure
D010743:Phospholipids