Abstract
Nitration of tyrosine residues in proteins is widely used as a marker for oxidative or nitrative stress in animal cells. Increase in tyrosine‐nitrated proteins with age in rat hippocampus and identification of several tyrosine‐nitrated proteins from aged rat muscles have been reported. However, no study on the protein nitration in the brain of adult‐ and middle‐aged rats has been demonstrated yet. In this study, we determined relative contents of nitrated proteins, nitric oxide synthases (NOS), and superoxide dismutases (SOD) in the hippocampus of adult‐ and middle‐aged F344 rats. We also determined difference in nitrated proteins between the adult‐ and middle‐aged rat hippocampus by using two‐dimensional electrophoresis (2DE). Western blot analyses showed that relative contents of nitrotyrosine‐containing proteins and expression of Mn‐SOD, EC‐SOD, nNOS, and eNOS were not different between the adult‐ and middle‐aged rats. However, the expression of Cu, Zn‐SOD was increased in the middle‐aged rat compared with that of adult rat. Furthermore, several spots for tyrosine‐nitrated protein in 2DE were different between the adult‐ and middle‐aged rats. Thus, although the nitrative and oxidative stress may not different so much between the adult‐ and middle‐aged rat hippocampus, target proteins for the nitration are changed. Proteome analysis of the nitrated proteins is underway in our laboratory.
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