Translocation of the 46 kDa protein(s) in response to activation of NADPH oxidase in guinea pig polymorphonuclear leukocytes.
Treatment of guinea pig polymorphonuclear leukocytes (PMNL) with phorbol 12-myristate 13-acetate (PMA) induced an increase in phosphorylation of 46 kDa protein(s) in parallel with activation of NADPH oxidase. In response to PMA stimulation, phosphorylated 46 kDa protein(s) increased markedly in the membrane fraction, accompanied by a decrease in the unphosphorylated form(s) in the cytosol. The results indicate that the 46 kDa protein(s) may be translocated concomitantly with its phosphorylation. On the other hand, in a cell-free activation system reconstituted from the cytosol and plasma membranes of unstimulated PMNL, arachidonic acid caused the translocation of the 46 kDa protein(s) from the cytosol to the plasma membranes concomitantly with an enhancement of NADPH oxidase activity. These results suggest that activation of NADPH oxidase is dependent on an association of 46 kDa protein(s) with the membranes both in intact PMNL and in the cell-free system.
Ohtsuka
T
T
Department of Physiological Chemistry, Hiroshima University School of Medicine.
Nakamura
M
M
Hiura
M
M
Yoshida
K
K
Okamura
N
N
Ishibashi
S
S
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000494:pharmacology; D001798:Blood Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005260:Female; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008970:Molecular Weight; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology*; D010449:Peptide Mapping; D010766:Phosphorylation; D012697:Serine Endopeptidases / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000596:Amino Acids; D001095:Arachidonic Acids; D001798:Blood Proteins; D016718:Arachidonic Acid; D009247:NADH, NADPH Oxidoreductases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D013755:Tetradecanoylphorbol Acetate