Requirements for extracellular metabolism of pulmonary surfactant: tentative identification of serine protease.
Pulmonary alveolar surfactant is secreted by the alveolar epithelium in the form of lamellar bodylike structures that evolve sequentially into tubular myelin and vesicular forms that can be separated by centrifugation. Using an in vitro procedure by which the extracellular metabolism of pulmonary surfactant can be mimicked, namely cyclic variation in surface area, we previously reported that serine protease activity, which we called "convertase," was required for the conversion of tubular myelin to the vesicular form. In the present studies we explored the biochemical requirements of this activity and sought the enzyme in alveolar products. Convertase activity has unusual requirements; in addition to being dependent on repetitive variations in surface area (cycling), it requires the presence of a high g fraction of lung secretions that is heat stable and not inhibitable by diisopropyl fluorophosphate (DFP) or alpha 1-antitrypsin, both typical serine protease inhibitors. The enzyme does not require calcium ions and has a pH optimum of 7.4. Convertase appears to be a component of surfactant itself because the ability of purified surfactant to convert to the vesicular form on cycling is impaired by pretreating it with DFP. A protein of Mr 75,000 that reacts with DFP and is heat sensitive was found in alveolar lavage, lamellar body preparations, and lung homogenate. It copurifies with lung surfactant in sucrose gradients. A similar DFP-reactive protein was observed in stable human neoplastic peripheral airway cell lines that express type II properties, suggesting that it may be a product of type II cells. We tentatively conclude that surfactant convertase is a 75,000 serine protease that is closely associated with surfactant phospholipid and that may be a product of alveolar type II cells.
Gross
N J
NJ
Department of Medicine, Stritch School of Medicine, Loyola University of Chicago, Illinois.
Schultz
R M
RM
D000818:Animals; D002118:Calcium / Q000378:metabolism; D004950:Esterases; D005110:Extracellular Space / Q000378:metabolism*; D005260:Female; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000378:metabolism; D051379:Mice; D010743:Phospholipids / Q000378:metabolism; D044345:Phosphoric Triester Hydrolases; D011663:Pulmonary Surfactants / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D014018:Tissue Distribution
D010743:Phospholipids; D011663:Pulmonary Surfactants; D006867:Hydrolases; D004950:Esterases; D044345:Phosphoric Triester Hydrolases; C020061:diisopropyl-fluorophosphatase; D012697:Serine Endopeptidases; D002118:Calcium