Calcium-dependent self-association of annexin II: a possible implication in exocytosis.
Alveolar type II cells secrete lung surfactant through exocytosis of lamellar bodies. We previously showed that the annexin II tetramer (Anx IIt) mediates the fusion of lamellar bodies with liposomes. The present study examined the possible involvement of membrane proteins in this process. Pre-treatment of lamellar bodies with trypsin and alpha-chymotrypsin reduced Anx IIt-mediated membrane fusion. With the use of an Anx IIt-conjugated Sepharose column, three Anx IIt-binding proteins with molecular weights of 67,000, 36,000 and 34,000 were isolated froM the Triton X-100 extract of bovine lung tissue membranes. These proteins were identified as annexins VI, II and IV by Western blot. The interaction of Anx IIt with annexins II and IV was confirmed by ligand blot assay. An EGTA-resistant membrane-bound annexin II was present in lung type II cells. Anx IIt preferentially hound to membranous annexin II compared with cytosolic annexin II of type II cells. With the use of immunofluorescence, annexin II was found to translocate from cytoplasm to plasma membranes in type II cells upon stimulation with phorbol 12-myristate 13-acetate. These results suggest that cytosolic annexin II may bind to membranous annexin II and form a protein-protein bridge to bring two membranes together.
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Department of Physiology, East Carolina University School of Medicine, Greenville, NC 27858, USA. lliu@brody.med.ecu.edu
D000818:Animals; D017306:Annexin A2 / Q000378:metabolism*; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002918:Chymotrypsin / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology*; D008024:Ligands; D008297:Male; D008561:Membrane Fusion / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014357:Trypsin / Q000378:metabolism
D017306:Annexin A2; D008024:Ligands; D002918:Chymotrypsin; D014357:Trypsin; D002118:Calcium