title: Annexin A7 trafficking to alveolar type II cell surface: possible roles for protein insertion into membranes and lamellar body secretion.
abstract: A role for annexin A7 (A7) is postulated in the obligatory fusion between lamellar bodies and the plasma membrane during surfactant secretion in alveolar type II cells. This study investigated if surfactant secretagogues increase cell surface A7, which could support A7 insertion into plasma membrane as annexin proteins reportedly lack membrane penetration ability. In vivo trafficking of A7 to cell surface was determined by immuno-staining after non-permeabilizing fixation of alveolar type II cells. Stimulation with various secretagogues increased protein kinase-dependent staining for A7 and ABCA3 in comparison to control cells. Biotin-labeling of surface proteins showed ~4% of total A7 in control cells, which increased ~3-4 folds in stimulated type II cells. Increased cell surface A7 was also observed by protein cross-linking studies showing ~70kDa A7-adduct in the membranes but not in the cytosol fraction of PMA- or A23187-stimulated cells. In vitro phosphorylation increased the Ca(2+)-dependent binding of recombinant A7 to lung plasma membranes; and subsequent cross-linking showed increased levels of ~70kDa A7-adduct. PMA-stimulation of type II cells increased A7 trafficking to lipid rafts suggesting that the latter are involved in A7 trafficking to the cell surface. However, in vitro membrane insertion of recombinant A7 and its tryptophan mutants as determined by fluorescence quenching with doxylPC suggested only shallow membrane insertion by A7. Together, our studies support in vivo association between surfactant secretion and cell surface A7 occurring by insertion into plasma membrane and by fusion of A7 containing lamellar bodies.
authors: Department of Pediatrics, Stony Brook University Medical Center, Stony Brook, NY, USA. Avinash.Jerath@sbumed.org
authors: Avinash
authors: A
authors: Chander
authors: Tudevdagva
authors: T
authors: Gerelsaikhan
authors: Pavan K
authors: PK
authors: Vasa
authors: Kelly
authors: K
authors: Holbrook
mesh_terms: D000818:Animals; D017310:Annexin A7 / Q000737:chemistry / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D008055:Lipids / Q000737:chemistry; D008168:Lung / Q000378:metabolism; D008561:Membrane Fusion / Q000187:drug effects; D021962:Membrane Microdomains / Q000737:chemistry; D008565:Membrane Proteins / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism; D011663:Pulmonary Surfactants / Q000378:metabolism*; D051381:Rats
chemical_list: D017310:Annexin A7; D008055:Lipids; D008565:Membrane Proteins; D011663:Pulmonary Surfactants