Comparison of spectrophotometric, HPLC and chemiluminescence methods for 3-nitrotyrosine and peroxynitrite interaction.
We have studied the interaction of 3-nitrotyrosine with peroxynitrite using three different methods; chemiluminescence, spectrophotometry and HPLC. Peroxynitrite-induced luminol or lucigenin chemiluminescence were significantly decreased by 3-nitrotyrosine, in concentration-dependent manners. The intensity of the peroxynitrite spectrum was also markedly reduced in the presence of 3-nitrotyrosine in the spectrophometric assay. However, there was no attenuation of the 3-nitrotyrosine signal in the HPLC assay after mixing with peroxynitrite. The interaction of 3-nitrotyrosine and hypochlorous acid (HOCl) was also studied via the chemiluminescence assay, where the HOCl-induced responses were markedly inhibited by 3-nitrotyrosine. These results suggest that caution should be taken when studying the levels or interactions of 3-nitrotyrosine.
Turan
Nilufer Nermin
NN
Gazi University, Faculty of Pharmacy, Department of Pharmacology, 06330 Etiler, Ankara, Turkey. niluferturan@gazi.edu.tr
Ark
Mustafa
M
Demiryurek
Abdullah Tuncay
AT
D002851:Chromatography, High Pressure Liquid; D006997:Hypochlorous Acid / Q000737:chemistry; D008163:Luminescent Measurements; D030421:Peroxynitrous Acid / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000737:chemistry*
D030421:Peroxynitrous Acid; C002744:3-nitrotyrosine; D014443:Tyrosine; D006997:Hypochlorous Acid