Synthesis of type II cell lamellar body lysozyme-15 kD protein (lbl-15) by perfused rat lung.
Isolated alveolar type II pneumocytes of the rat have been shown to secrete a 14 to 15 kD protein that has some sequence homology and immunoreactivity with lysozyme. Using immunochemical analyses of rat lung subcellular fractions and 35S metabolic labeling of isolated perfused lung preparations, we studied the subcellular distribution and synthetic pathway for this protein. SDS-PAGE and Western blotting of lamellar bodies (LB) using a polyclonal anti-human lysozyme (anti-HLZ) demonstrated a single band at 15 kD that was significantly enriched over rat lung homogenates, isolated lysosomes, and type II cell lysates. This 15 kD protein isolated from LB by immunoprecipitation with anti-HLZ also demonstrated functional lysozyme activity and was termed lamellar body lysozyme (lbl-15). Analysis of LB and surfactant (SF) isolated from 12 separate perfused lung preparations labeled for 6 h with 35S-labeled amino acids demonstrated that lbl-15 represented a significant portion of the radiolabeled LB proteins (5.9% of total LB radioactivity). Lamellar bodies and an extracellular fraction (surfactant) obtained from rat lungs perfused with 35S-methionine-cysteine for varying times both showed time-dependent appearance of lbl-15. At all time points, the specific activity of lbl-15 was greater in LB than in SF. The kinetics for appearance of lbl-15 in LB and SF was similar to that for surfactant protein A. These results indicate that in the rat lung, type II cells synthesize a 15 kD protein (lbl-15) that is secreted into the alveolar space via an organellar pathway involving LB.(ABSTRACT TRUNCATED AT 250 WORDS)
Beers
M F
MF
Institute for Environmental Medicine University of Pennsylvania School of Medicine, Philadelphia 19104-6068.
Kim
C Y
CY
Dodia
C
C
Fisher
A B
AB
D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007150:Immunohistochemistry; D007700:Kinetics; D008168:Lung / Q000201:enzymology*; D008970:Molecular Weight; D009113:Muramidase / Q000032:analysis / Q000096:biosynthesis*; D015388:Organelles / Q000201:enzymology*; D010477:Perfusion; D011510:Proteolipids / Q000096:biosynthesis; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000096:biosynthesis; D011865:Radioisotope Dilution Technique; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology; D013462:Sulfur Radioisotopes
D000906:Antibodies; D011510:Proteolipids; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D013462:Sulfur Radioisotopes; C079018:LBL-15 protein, rat; D009113:Muramidase