期刊
PLANTA
卷 236, 期 2, 页码 427-436出版社
SPRINGER
DOI: 10.1007/s00425-012-1614-1
关键词
Acylamino acid-releasing enzyme; Antioxidative system; Oxidative stress; Oxidized protein hydrolase; Protein modification
资金
- Global Center of Excellence for Dryland Science of the Arid Land Research Center, Tottori University
- Nara Institute of Science and Technology
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- Grants-in-Aid for Scientific Research [11F01100] Funding Source: KAKEN
Acylamino acid-releasing enzyme/oxidized protein hydrolase (AARE/OPH) has been biochemically demonstrated to be a bifunctional protease that has exopeptidase activity against N-alpha-acylated peptides and endopeptidase activity against oxidized and glycated proteins; however, its physiological role remains unknown. In this study, to determine its physiological significance, we produced AARE/OPH-overexpressing and -suppressed plants and assessed the biological impacts of AARE/OPH. The subcellular localization of Arabidopsis AARE/OPH was found to be cytoplasmic and nuclear by transient expression analysis of tdTomato-fused Arabidopsis AARE/OPH. Overexpression of AARE/OPH exhibited no apparent effect on the level of oxidized proteins because wild types probably have inherently high AARE/OPH activity. Through RNAi gene suppressing, we successfully produced AARE/OPH-suppressed Arabidopsis plants (aare) that exhibited almost no AARE activity. In the aare plant, electrolyte leakage by methyl viologen treatment was enhanced compared to that of non-transformant plants, suggesting that the plasma membranes of aare easily suffered oxidative damage, probably as a result of deterioration of the cytoplasmic antioxidative system. Correspondingly, proteomic analysis revealed that the aare plant accumulated a number of oxidized proteins including cytoplasmic antioxidant enzymes. On the basis of these results, we concluded that AARE/OPH plays a homeostatic role in sustaining the cytoplasmic antioxidative system.
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