Phox activity of differentiated PLB-985 cells is enhanced, in an agonist specific manner, by the PLA2 activity of Prdx6-PLA2.
Peroxiredoxin 6-phospholipase A(2)  (Prdx6-PLA(2) ) is a bi-functional enzyme with peroxi-redoxin (Prdx) and phospholipase A(2)  (PLA(2) ) activities. To investigate its impact on phagocyte NADPH oxidase (phox) activity in a neutrophil model, the protein was knocked down in PLB-985 cells using stable expression of a small hairpin RNA (shRNA) and phox activity was monitored after cell differentiation. The knockdown cells had reduced oxidase activity in response to stimulation with the formylated peptide fMLF, but the response to the phorbol ester PMA was unchanged. Reintroduction of shRNA-resistant Prdx6-PLA(2)  into the knockdown cells by stable transfection with a Prdx6-PLA(2)  expression plasmid restored the fMLF response, as did reintroduction of Prdx6-PLA(2)  mutated in the Prdx active site; reintroduction of PLA(2)  active site mutants, however, failed to restore the response. Thus, the PLA(2)  activity of Prdx6-PLA(2)  in intact cells mediates its ability to enhance phox activity in response to fMLF. In combination with previous publications by other groups, our work indicates that various PLA(2)  isoforms can enhance oxidase activity but they are differentially important in different cell types and in the response to different agonists.
Bonfils Blood Center, Denver, CO 80230, USA.
Michael A
MA
Ellison
Gail W
GW
Thurman
Daniel R
DR
Ambruso
D045744:Cell Line, Tumor; D006801:Humans; D007951:Leukemia, Myeloid / Q000201:enzymology*; D019255:NADPH Oxidases / Q000378:metabolism*; D054465:Peroxiredoxin VI / Q000378:metabolism*; D054467:Phospholipases A2 / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
D013481:Superoxides; D014212:Tretinoin; C515977:PRDX6 protein, human; D054465:Peroxiredoxin VI; D019255:NADPH Oxidases; D054467:Phospholipases A2; D013755:Tetradecanoylphorbol Acetate