Cadmium causes the oxidative modification of proteins in pea plants
Cadmium causes the oxidative modification of proteins in pea plants
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DOI：
10.1046/j.1365-3040.2002.00850.x
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发表时间：
2002-05-01
影响因子：
7.3
通讯作者：
Sandalio, LM
中科院分区：
文献类型：
--
作者：
Romero-Puertas, MC;Palma, JM;Sandalio, LM
豌豆（Pisum sativum L.）研究了在50 μ M CdCl 2存在下生长的植物叶片中蛋白质中羰基的氧化产生、蛋白质降解速率和蛋白水解活性。在叶提取物中的羰基含量测量衍生与2，4-二硝基苯肼（DNPH），是两倍高，在植物处理镉比对照植物。氧化蛋白的鉴定通过DNPH衍生的蛋白质的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和抗DNPH抗体的免疫化学检测进行。镉暴露的植物中的反应带的强度高于对照。通过使用不同的抗体，一些氧化蛋白被确定为Rubisco，谷胱甘肽还原酶，锰超氧化物歧化酶，和过氧化氢酶。叶粗提物与H2 O2浓度增加的孵育表现出逐步增强羰基含量和氧化蛋白的模式是类似的，发现在镉处理的植物。氧化的蛋白质被更有效地降解，并且由于金属处理，蛋白水解活性增加了20%。在过氧化物酶体纯化豌豆叶中的羰基含量的上升类似于从镉处理的植物粗提取物中得到的观察，但过氧化物酶体膜的功能没有明显的影响镉。所获得的结果表明参与的氧化应激，可能介导的H2 O2，和蛋白水解降解的机制中的镉毒性豌豆植物的叶子，他们似乎参与了镉诱导的衰老以前报道在这些植物。
In pea (Pisum sativum L.) leaves from plants grown in the presence of 50 muM CdCl2 the oxidative production of carbonyl groups in proteins, the rate of protein degradation and the proteolytic activity were investigated. In leaf extracts the content of carbonyl groups measure derivatization with 2,4-dinitrophenylhydrazine (DNPH), was two-fold higher in plants treated with Cd than in control plants. The identification of oxidized proteins was carried out by sodium dodecyl sulphate-polyacrylamide gel electrophoresis of proteins derivatized with DNPH and immunochemical detection with an antibody against DNPH. The intensity of the reactive bands was higher in plants exposed to Cd than in controls. By using different antibodies some of the oxidized proteins were identified as Rubisco, glutathione reductase, manganese superoxide dismutase, and catalase. The incubation of leaf crude extracts with increasing H2O2 concentrations showed a progressive enhancement in carbonyl content and the pattern of oxidized proteins was similar to that found in Cd-treated plants. Oxidized proteins were more efficiently degraded, and the proteolytic activity increased 20% due to the metal treatment. In peroxisomes purified from pea leaves a rise in the carbonyl content similar to that obtained in crude extracts from Cd-treated plants was observed, but the functionality of the peroxisomal membrane was not apparently affected by Cd. Results obtained demonstrate the participation of both oxidative stress, probably mediated by H2O2, and proteolytic degradation in the mechanism of Cd toxicity in leaves of pea plants, and they appear to be involved in the Cd-induced senescence previously reported in these plants.