Transient translocation and activation of protein phosphatase 2A during mast cell secretion.
Okadaic acid inhibits secretion from mast cells, suggesting a regulatory role for protein Ser/Thr phosphatases type I (PP1) and/or 2A (PP2A) in the secretory process. In unstimulated RBL-2H3 cells, okadaic acid pretreatment inhibited PP2A activity in both cytosol and membrane fractions, but inhibition of secretion correlated with inhibition of membrane-bound rather than cytosolic PP2A activity. Okadaic acid had very little effect on PP1 activity. Stimulation of RBL-2H3 cells by antigen led to the activity and amount of PP2A in the membrane fraction increasing nearly 2-fold. In contrast, there was little change in the activity or distribution of PP1. Importantly, the translocation of PP2A was transient, coinciding with or marginally preceding the peak rate of secretion, suggesting a link between PP2A translocation, activity, and secretion. Phorbol 12-myristate 13-acetate plus the calcium ionophore A23187 induced a slower, prolonged rate of secretion that coincided with a similarly protracted translocation of PP2A to the membrane fraction. PP2A translocation is not the only event required for secretion as translocation was also induced by phorbol 12-myristate 13-acetate, without resulting in secretion. These results indicate that increased protein dephosphorylation in the membrane fraction mediated by PP2A is required for mast cell secretion. To our knowledge, this is the first demonstration of a signal-mediated, rapid, transient translocation and activation of PP2A in membranes in any system.
Ludowyke
R I
RI
Centre for Immunology, St. Vincent's Hospital, University of New South Wales, Sydney, New South Wales 2010, Australia. r.ludowyke@cfi.unsw.edu.au
Holst
J
J
Mudge
L M
LM
Sim
A T
AT
D000818:Animals; D000941:Antigens / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D004140:Dinitrophenols / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D007073:Immunoglobulin E / Q000378:metabolism; D008387:Marine Toxins; D008407:Mast Cells / Q000201:enzymology* / Q000378:metabolism; D019319:Okadaic Acid / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D054648:Protein Phosphatase 2; D051381:Rats; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D000941:Antigens; D004140:Dinitrophenols; D008387:Marine Toxins; D010080:Oxazoles; C034638:dinitrophenyl-bovine serum albumin; D019319:Okadaic Acid; D012710:Serum Albumin, Bovine; D007073:Immunoglobulin E; D000001:Calcimycin; C059041:calyculin A; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D001619:beta-N-Acetylhexosaminidases; D013755:Tetradecanoylphorbol Acetate