Evidence for participation of vicinal dithiols in the activation sequence of the respiratory burst of human neutrophils.
Phenylarsine oxide (PAO) specifically forms a stable ring complex with vicinal dithiols that can be reversed with 2,3-dimercaptopropanol (DMP). Pretreatment of human neutrophils with micromolar concentrations of PAO inhibited release of superoxide anion (O2-) stimulated by N-formyl-methionyl-leucyl-phenylalanine (FMLP) or phorbol 12-myristate 13-acetate (PMA); the inhibition was reversed with DMP, but not with 2-mercaptoethanol. PAO did not affect O2- release in previously stimulated cells. PAO did not affect the FMLP-induced Ca2+ response, suggesting that PAO affects a postreceptor event that does not modulate the Ca2+ transient. Treatment of isolated membrane or cytosolic fractions with PAO did not change the rates of arachidonate-stimulated O2- production in a cell-free system. Pretreatment of unstimulated neutrophils with PAO inactivated cytosolic protein kinase C (PKC); the inactivation was reversed with DMP. However, PAO did not affect PMA-induced translocation of beta-PKC protein or reduce the PKC activity translocated to the membrane. PAO had no effect on tyrosine kinase activity but inactivated phosphotyrosine phosphatase; stimulus-induced tyrosine phosphorylation of several proteins was markedly enhanced. These results suggest that vicinal dithiols play an essential role in activation of the respiratory burst oxidase. Possible sites for the activity of these essential vicinal dithiols include PKC and the regulatory balance of tyrosine phosphatase activity and tyrosine phosphorylation.
Kutsumi
H
H
Department of Preventive Medicine, Kyoto Prefectural University of Medicine, Japan.
Kawai
K
K
Johnston
R B
RB
Rokutan
K
K
D016718:Arachidonic Acid / Q000494:pharmacology; D001152:Arsenicals / Q000494:pharmacology*; D001798:Blood Proteins / Q000378:metabolism; D002118:Calcium / Q000097:blood; D002474:Cell-Free System; D003600:Cytosol / Q000187:drug effects; D004112:Dimercaprol / Q000494:pharmacology; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000097:blood; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000097:blood; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D016897:Respiratory Burst / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D013438:Sulfhydryl Compounds / Q000097:blood*; D013481:Superoxides / Q000097:blood; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D001152:Arsenicals; D001798:Blood Proteins; D007527:Isoenzymes; D013438:Sulfhydryl Compounds; D004112:Dimercaprol; C029341:oxophenylarsine; D013481:Superoxides; D016718:Arachidonic Acid; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium