In vitro fusion of lung lamellar bodies and plasma membrane is augmented by lung synexin.
Lamellar bodies of lung epithelial type II cells undergo fusion with plasma membrane prior to exocytosis of surfactant into the alveolar lumen. Since synexin from adrenal glands promotes aggregation and fusion of chromaffin granules, we purified synexin-like proteins from bovine lung cytosolic fraction, and evaluated their effect on the fusion of isolated lamellar bodies and plasma membrane fractions. Synexin activity, which co-purified with an approx. 47 kDa protein (pI 6.8), was assessed by following calcium-dependent aggregation of liposomes prepared from a mixture of phosphatidylcholine:phosphatidylserine (PC:PS, 3:1, mol/mol). Lung synexin caused aggregation of liposomes approximating lung surfactant lipid-like composition, isolated lamellar bodies, or isolated plasma membrane fraction. Lung synexin promoted fusion only in the presence of calcium. It augmented fusion between lamellar bodies and plasma membranes, lamellar bodies and liposomes, or between two populations of liposomes. However, selectivity with regard to synexin-mediated fusion was observed as synexin did not promote fusion between plasma membrane and liposomes, or between liposomes of surfactant lipid-like composition and other liposomes. These observations support a role for lung synexin in membrane fusion between the plasma membrane and lamellar bodies during exocytosis of lung surfactant, and suggest that such fusion is dependent on composition of interacting membranes.
Chander
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Institute for Environmental Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104.
Wu
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D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D017310:Annexin A7; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002462:Cell Membrane / Q000502:physiology*; D004848:Epithelium / Q000648:ultrastructure; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008168:Lung / Q000737:chemistry / Q000648:ultrastructure*; D008297:Male; D008561:Membrane Fusion / Q000187:drug effects*; D008969:Molecular Sequence Data; D015388:Organelles / Q000502:physiology*; D010713:Phosphatidylcholines / Q000032:analysis; D010718:Phosphatidylserines / Q000032:analysis; D011506:Proteins / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains
D000596:Amino Acids; D017310:Annexin A7; D008081:Liposomes; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D011506:Proteins; D002118:Calcium