Nitric oxide activation of TrkB through peroxynitrite.
NIH-3T3 cells stably transfected with TrkB, the receptor for brain-derived neurotrophic factor (BDNF), were used to study the effects of NO and peroxynitrite on TrkB. 3-Morpholinosydnonimine (SIN-1), a donor of NO and O2- which immediately react to form peroxynitrite, induced TrkB tyrosine phosphorylation in a dose-dependent relationship from 2 to 40 mM. TrkB phosphorylation by SIN-1 was blocked by superoxide dismutase, which converts O2 to H2O2 and prevents its reaction with NO to form peroxynitrite, and by K252a, an inhibitor of TrkB phosphorylation by BDNF. Treatment with NO or O2- alone did not activate TrkB. Treatment directly with 1-4 mM peroxynitrite resulted in a dose-dependent increase in tyrosine phosphorylation of TrkB. SIN-1 treatment induced tyrosine phosphorylation of phospholipase C-gamma1 (PLC-gamma1) and induced its binding with activated TrkB, similar to that seen with BDNF downstream signaling pathways. These studies demonstrate activation of TrkB through peroxynitrite.
Yuen
E C
EC
Department of Neurology, University of Washington, Seattle 98195, USA.
Gunther
E C
EC
Bothwell
M
M
D016475:3T3 Cells; D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D006801:Humans; D026121:Indole Alkaloids; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D008981:Molsidomine / Q000031:analogs & derivatives / Q000494:pharmacology; D009566:Nitrates / Q000494:pharmacology*; D009569:Nitric Oxide / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D016877:Oxidants / Q000494:pharmacology; D051966:Phospholipase C gamma; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D051381:Rats; D020813:Receptor, trkB / Q000187:drug effects / Q000502:physiology*; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000494:pharmacology; D014162:Transfection; D010738:Type C Phospholipases / Q000378:metabolism
D019208:Brain-Derived Neurotrophic Factor; D002227:Carbazoles; D026121:Indole Alkaloids; D007527:Isoenzymes; D009566:Nitrates; D020030:Nitric Oxide Donors; D016877:Oxidants; D011994:Recombinant Proteins; D019000:Phosphotyrosine; C039642:peroxynitric acid; D009569:Nitric Oxide; C002385:linsidomine; C049985:staurosporine aglycone; D008981:Molsidomine; D013482:Superoxide Dismutase; D020813:Receptor, trkB; D010738:Type C Phospholipases; D051966:Phospholipase C gamma