title: Calcium ionophore and phorbol ester increase membrane binding of annexin a7 in alveolar type II cells.
abstract: The fusion of lamellar body with plasma membrane, a distal obligatory step in exocytosis of lung surfactant, may be mediated by annexin a7 (anx a7; synexin). To understand the mechanism of anx a7 action, we tested the hypothesis that anx a7 binding to membranes would increase in order to facilitate membrane fusion during stimulation of lung surfactant secretion. Isolated rat alveolar type II cells were treated with established secretagogues of lung surfactant and the membrane and cytosol fractions were analyzed for in vitro binding of anx a7. In cells treated with calcium ionophore (A23187) or phorbol 12-myristate 13-acetate (PMA), anx a7 binding to the membrane fraction was increased by 120%, while that to the cytosol fraction was decreased by 40%, when compared with binding to corresponding fractions from control cells. Protein kinase inhibitors prevented the PMA effects on anx a7 binding. The lamellar body and plasma membrane fractions of A23187-treated cells also showed increased binding of anx a7. The lamellar bodies of A23187-treated cells showed lower K(m) for Ca(2+) and higher maximal binding of anx a7, when compared with those from control cells. Collectively, our findings suggest that these two agents modify membrane proteins to regulate anx a7 binding, which may facilitate increased membrane fusion activity during stimulation of surfactant secretion.
authors.affiliation: Department of Pediatrics, Division of Neonatology, State University of New York, Stony Brook, NY 11794-8111, USA. achander@mail.som.sunysb.edu
authors.forename: Avinash
authors.initials: A
authors.lastname: Chander
authors.forename: Namita
authors.initials: N
authors.lastname: Sen
authors.forename: Devendra G
authors.initials: DG
authors.lastname: Naidu
authors.forename: Alan R
authors.initials: AR
authors.lastname: Spitzer
mesh_terms: D000818:Animals; D017310:Annexin A7 / Q000378:metabolism*; D012634:Bodily Secretions / Q000187:drug effects / Q000502:physiology; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D008561:Membrane Fusion / Q000187:drug effects / Q000502:physiology; D010703:Phorbol Esters / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D011650:Pulmonary Alveoli / Q000187:drug effects / Q000378:metabolism*; D011663:Pulmonary Surfactants / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
chemical_list: D017310:Annexin A7; D007476:Ionophores; D010703:Phorbol Esters; D011663:Pulmonary Surfactants; D002118:Calcium