Pervanadate activates NADPH oxidase via protein kinase C-independent phosphorylation of p47-phox.
We studied differences between the NADPH oxidase activation pathways triggered by pervanadate, a protein tyrosine phosphatase inhibitor, and phorbol 12-myristate 13-acetate (PMA), a protein kinase C activator, in guinea pig neutrophils. Previously, pervanadate has been shown to activate NADPH oxidase via the tyrosine kinase-dependent pathway (Yamaguchi et al. Arch. Biochem. Biophys. 323, 382-386, 1995). Both pervanadate and PMA induced superoxide anion (O-2) production, translocation of the 47-kDa protein component of the phagocyte oxidase (p47-phox) to the plasma membrane, and phosphorylation of p47-phox in the membrane. A selective protein kinase C inhibitor, GF 109203X, markedly inhibited PMA-induced O-2 production, p47-phox translocation, and p47-phox phosphorylation, but did not inhibit pervanadate-induced O-2 production and only slightly suppressed pervanadate-induced translocation and phosphorylation. These results demonstrate that pervanadate activates NADPH oxidase independently of protein kinase C. Phosphopeptide mapping of p47-phox revealed differences in the mechanism between pervanadate-induced and PMA-induced phosphorylation. Furthermore, some protein kinases which phosphorylate p47-phox-derived peptide are activated by pervanadate. These results suggest the existence of novel protein kinases responsible for the phosphorylation of p47-phox and the activation of these protein kinases by tyrosine kinase.
Yaname
H
H
Department of Physiological Chemistry, Hiroshima University School of Medicine, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8551, Japan.
Fukunaga
T
T
Nigorikawa
K
K
Okamura
N
N
Ishibashi
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D001692:Biological Transport / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D006168:Guinea Pigs; D007211:Indoles / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D009252:NADPH Dehydrogenase / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors; D013481:Superoxides / Q000037:antagonists & inhibitors / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology*
D007211:Indoles; D008301:Maleimides; D010750:Phosphoproteins; C076715:pervanadate; D013481:Superoxides; D014638:Vanadates; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; D009252:NADPH Dehydrogenase; D011494:Protein Kinases; D011493:Protein Kinase C; D017027:Protein Tyrosine Phosphatases; C070515:bisindolylmaleimide I; D013755:Tetradecanoylphorbol Acetate