Differential abilities of nitrogen dioxide and nitrite to nitrate proteins in thylakoid membranes isolated from Arabidopsis leaves.
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.
a Department of Mathematical and Life Sciences , Graduate School of Science, Hiroshima University , Higashi-Hiroshima, Hiroshima , Japan.
Misa
M
Takahashi
a Department of Mathematical and Life Sciences , Graduate School of Science, Hiroshima University , Higashi-Hiroshima, Hiroshima , Japan.
Jun
J
Shigeto
a Department of Mathematical and Life Sciences , Graduate School of Science, Hiroshima University , Higashi-Hiroshima, Hiroshima , Japan.
Tatsuo
T
Shibata
a Department of Mathematical and Life Sciences , Graduate School of Science, Hiroshima University , Higashi-Hiroshima, Hiroshima , Japan.
Atsushi
A
Sakamoto
a Department of Mathematical and Life Sciences , Graduate School of Science, Hiroshima University , Higashi-Hiroshima, Hiroshima , Japan.
Hiromichi
H
Morikawa
D017360:Arabidopsis / Q000187:drug effects / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D009573:Nitrites / Q000494:pharmacology*; D009585:Nitrogen Dioxide / Q000494:pharmacology*; D018515:Plant Leaves / Q000187:drug effects / Q000378:metabolism*; D020524:Thylakoids / Q000187:drug effects / Q000378:metabolism*
D029681:Arabidopsis Proteins; D009573:Nitrites; D009585:Nitrogen Dioxide
Arabidopsis; PsbO; nitrite; nitrogen dioxide; protein tyrosine nitration; thylakoid membranes
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