Phosphorylation by protein kinase C of annexin 2 in chromaffin cells stimulated by nicotine.
Annexin 2 phosphorylated in vitro by protein kinase C has been shown to restore partially catecholamine secretion in streptolysin O-permeabilized chromaffin cells depleted of their protein kinase C activity. This result suggested a phosphorylation of annexin 2 in stimulated cells. Nicotine stimulation induced an increase of 32P incorporation in annexin 2 heavy chain concomitant with catecholamine release. This incorporation results from phosphorylation by protein kinase C because (a) serine was the only phosphorylated residue, (b) 32P incorporation was inhibited by the protein kinase inhibitors H7, GF 109203X, and staurosporine, and (c) activators of this enzyme, 12-O-tetradecanoylphorbol 13-acetate and 1,2-dioctanoylglycerate, increased the incorporation of radioactivity. The phosphorylated heavy chain had an electrophoretic mobility lower than that of the unmodified one, thus allowing determination of the fraction of phosphorylated protein. In the resting state, a significant fraction of annexin 2 heavy chain was phosphorylated, and nicotine stimulation resulted in an activation of both phosphorylation and dephosphorylation. Phosphorylation was largely increased in the presence of okadaic acid, indicating the involvement of type 1 and 2A phosphatases.
Delouche
B
B
Centre National de la Recherche Scientifique, UPR 9071, Neurobiologie Physico-Chimique, Institut de Biologie Physico-Chimique, Paris, France.
Pradel
L A
LA
Henry
J P
JP
D000818:Animals; D017306:Annexin A2 / Q000378:metabolism*; D001426:Bacterial Proteins; D002395:Catecholamines / Q000378:metabolism; D002417:Cattle; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects / Q000201:enzymology; D019439:Chromaffin Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D007399:Interphase / Q000502:physiology; D008301:Maleimides / Q000494:pharmacology; D009538:Nicotine / Q000494:pharmacology*; D018722:Nicotinic Agonists / Q000494:pharmacology*; D010710:Phosphates / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology; D013301:Streptolysins / Q000494:pharmacology
D017306:Annexin A2; D001426:Bacterial Proteins; D002395:Catecholamines; D004791:Enzyme Inhibitors; D007211:Indoles; D008301:Maleimides; D018722:Nicotinic Agonists; D010710:Phosphates; D010761:Phosphorus Radioisotopes; D013301:Streptolysins; C025964:streptolysin O; D009538:Nicotine; D011493:Protein Kinase C; D019311:Staurosporine; C070515:bisindolylmaleimide I