biostudies-literatureUnknown11(10)Takahashi MExposure of Arabidopsis leaves to nitrogen dioxide (NO<sub>2</sub>) results in nitration of specific chloroplast proteins. To determine whether NO<sub>2</sub> itself and/or nitrite derived from NO<sub>2</sub> can nitrate proteins, Arabidopsis thylakoid membranes were isolated and treated with NO<sub>2</sub>-bubbled or potassium nitrite (KNO<sub>2</sub>) buffer, followed by protein extraction, electrophoresis, and immunoblotting using an anti-3-nitrotyrosine (NT) antibody. NO<sub>2</sub> concentrations in the NO<sub>2</sub>-bubbled buffer were calculated by numerically solving NO<sub>2</sub> dissociation kinetic equations. The two buffers were adjusted to have identical nitrite concentrations. Both treatments yielded an NT-immunopositive band that LC/MS identified as PSBO1. The difference in the band intensity between the 2 treatments was designated nitration by NO<sub>2</sub>. Both NO<sub>2</sub> and nitrite mediated nitration of proteins, and the nitration ability per unit NO<sub>2</sub> concentration was ∼100-fold greater than that of nitrite.Plant signaling & behaviore1237329https://www.ebi.ac.uk/biostudies/studies/S-EPMC5117089biostudies-literatureDifferential abilities of nitrogen dioxide and nitrite to nitrate proteins in thylakoid membranes isolated from Arabidopsis leaves.PMC5117089Shibata TShigeto JSakamoto ATakahashi MMorikawa HfalseDifferential abilities of nitrogen dioxide and nitrite to nitrate proteins in thylakoid membranes isolated from Arabidopsis leaves.Exposure of Arabidopsis leaves to nitrogen dioxide (NO<sub>2</sub>) results in nitration of specific chloroplast proteins. To determine whether NO<sub>2</sub> itself and/or nitrite derived from NO<sub>2</sub> can nitrate proteins, Arabidopsis thylakoid membranes were isolated and treated with NO<sub>2</sub>-bubbled or potassium nitrite (KNO<sub>2</sub>) buffer, followed by protein extraction, electrophoresis, and immunoblotting using an anti-3-nitrotyrosine (NT) antibody. NO<sub>2</sub> concentrations in the NO<sub>2</sub>-bubbled buffer were calculated by numerically solving NO<sub>2</sub> dissociation kinetic equations. The two buffers were adjusted to have identical nitrite concentrations. Both treatments yielded an NT-immunopositive band that LC/MS identified as PSBO1. The difference in the band intensity between the 2 treatments was designated nitration by NO<sub>2</sub>. Both NO<sub>2</sub> and nitrite mediated nitration of proteins, and the nitration ability per unit NO<sub>2</sub> concentration was ∼100-fold greater than that of nitrite.2016-01-01T00:00:00Z2016 Oct2025-04-19T03:03:58.148Z2019-03-27T02:29:25ZS-EPMC51170892766177110.1080/15592324.2016.1237329