A possible biochemical link between NADPH oxidase (Nox) 1 redox-signalling and ERp72
A possible biochemical link between NADPH oxidase (Nox) 1 redox-signalling and ERp72
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DOI：
10.1042/bj20071259
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发表时间：
2008-11-15
影响因子：
4.1
通讯作者：
Kamata, Tohru
中科院分区：
文献类型：
--
作者：
Chen, Wei;Shang, Wei Hao;Kamata, Tohru
新出现的证据表明，Nox（NADPH氧化酶）1产生的ROS（活性氧）在各种细胞过程中发挥关键的调节作用，但很少有人知道的氧化酶的直接目标。在本研究中，我们表明，选择性氧化的蛋白质响应Nox 1产生的ROS是ERp72（内质网蛋白72 kDa）与TRX（硫氧还蛋白）同源结构域。Nox 1对ERp72的氧化导致其还原酶活性的抑制。EGF处理的细胞刺激的Nox1活性和激活的Nox1随后介导的EGF诱导的ERp72还原酶活性的抑制。免疫共沉淀、GST（谷胱甘肽转移酶）pulldown分析和突变分析表明，Nox 1与ERp72相关，其N端包含一个Ca 2+结合位点和第一个TRX样基序。此外，共聚焦显微镜显示Nox 1和ERp72在质膜上的共定位。这些结果表明，Nox 1与ERp72功能相关，在细胞环境中调节氧化还原敏感的信号通路。
Emerging evidence indicates that Nox (NADPH oxidase) 1-generated ROS (reactive oxygen species) play critical regulatory roles in various cellular processes, yet little is known of direct targets for the oxidase. In the present study we show that one of the proteins selectively oxidized in response to Nox 1-generated ROS was ERp72 (endoplasmic reticulum protein 72 kDa) with TRX (thioredoxin) homology domains. Oxidation of ERp72 by Nox1 resulted in an inhibition of its reductase activity. EGF treatment of cells stimulated the Nox1 activity and the activated Nox1 subsequently mediated EGF-induced suppression of the ERp72 reductase activity. Co-immunoprecipitation, GST (glutathione transferase) pulldown assays and mutational analysis, indicated that Nox1 associates with ERp72, which involves its N-tenninus encompassing a Ca2+-binding site and the first TRX-like motif. Furthermore, confocal microscopy showed co-localization between Nox1 and ERp72 at the plasma membrane. These results suggest that Nox 1 functionally associates with ERp72, regulating redox-sensitive signalling pathways in a cellular context.