Differential abilities of nitrogen dioxide and nitrite to nitrate proteins in thylakoid membranes isolated from Arabidopsis leaves

Misa Takahashi, Jun Shigeto, Tatsuo Shibata, Atsushi Sakamoto, Hiromichi Morikawa

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

Exposure of Arabidopsis leaves to nitrogen dioxide (NO2) results in nitration of specific chloroplast proteins. To determine whether NO2 itself and/or nitrite derived from NO2 can 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. NO2 concentrations in the NO2-bubbled buffer were calculated by numerically solving NO2 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 NO2. Both NO2 and nitrite mediated nitration of proteins, and the nitration ability per unit NO2 concentration was ∼100-fold greater than that of nitrite.

Original languageEnglish
Article numbere1237329
JournalPlant Signaling and Behavior
Volume11
Issue number10
DOIs
Publication statusPublished - Oct 2 2016

All Science Journal Classification (ASJC) codes

  • Plant Science

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