Differential abilities of nitrogen dioxide and nitrite to nitrate proteins in thylakoid membranes isolated from Arabidopsis leaves
Differential abilities of nitrogen dioxide and nitrite to nitrate proteins in thylakoid membranes isolated from Arabidopsis leaves
复制标题
二氧化氮和亚硝酸盐对拟南芥叶中分离的类囊体膜中硝酸盐蛋白质的差异能力
DOI：
10.1080/15592324.2016.1237329
复制
发表时间：
2016
影响因子：
2.9
通讯作者：
Morikawa H
中科院分区：
文献类型：
--
作者：
Takahashi M;Shigeto J;Shibata T;Sakamoto A;Morikawa H
拟南芥叶片暴露于二氧化氮（NO2）的结果在特定的叶绿体蛋白质硝化。为了确定是否NO2本身和/或来自NO2的亚硝酸盐可以硝酸蛋白质，分离拟南芥类囊体膜，并用NO2-鼓泡或亚硝酸钾（KNO 2）缓冲液处理，然后进行蛋白质提取，电泳和免疫印迹使用抗3-硝基酪氨酸（NT）抗体。通过数值求解NO2解离动力学方程，计算了NO2-鼓泡缓冲液中NO2的浓度。将两种缓冲液调节至具有相同的亚硝酸盐浓度。两种处理均产生NT-免疫阳性条带，LC/MS鉴定为PSBO 1。2个处理之间的谱带强度的差异被指定为NO2硝化。NO2和亚硝酸盐都能介导蛋白质的硝化，单位NO2浓度的硝化能力是亚硝酸盐的100倍。
Exposure of Arabidopsis leaves to nitrogen dioxide (NO2) results in nitration of specific chloroplast proteins. To determine whether NO2itself and/or nitrite derived from NO2can nitrate proteins, Arabidopsis thylakoid membranes were isolated and treated with NO2-bubbled or potassium nitrite (KNO2) buffer, followed by protein extraction, electrophoresis, and immunoblotting using an anti-3-nitrotyrosine (NT) antibody. NO2concentrations in the NO2-bubbled buffer were calculated by numerically solving NO2dissociation 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 NO2. Both NO2and nitrite mediated nitration of proteins, and the nitration ability per unit NO2concentration was ∼100-fold greater than that of nitrite.