S-nitrosylated proteins of a medicinal CAM plant Kalanchoe pinnata -: ribulose-1,5-bisphosphate carboxylase/oxygenase activity targeted for inhibition
S-nitrosylated proteins of a medicinal CAM plant Kalanchoe pinnata -: ribulose-1,5-bisphosphate carboxylase/oxygenase activity targeted for inhibition
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DOI：
10.1111/j.1742-4658.2008.06425.x
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发表时间：
2008-06-01
期刊：
影响因子：
5.4
通讯作者：
Deswal, Renu
中科院分区：
文献类型：
--
作者：
Abat, Jasmeet K.;Mattoo, Autar K.;Deswal, Renu
一氧化氮（NO）是一种影响植物无数过程的信号分子。然而，机械细节是有限的。NO翻译后通过半胱氨酸的s -亚硝基化修饰蛋白质。采用生物素开关技术对药用植物天冬氨酸酸代谢（CAM）植物凤尾莲（kalanche pinnata）的可溶性s -亚硝基蛋白质组进行了纯化。MALDI-TOF质谱鉴定了19个靶点，包括与碳、氮和硫代谢、细胞骨架、胁迫和光合作用相关的蛋白质。其中一些与先前在拟南芥中发现的相似，但激酶样蛋白、乙醇酸氧化酶、推定的UDP葡萄糖4-外聚酶和推定的DNA拓扑异构酶II尚未被确定为任何生物体的靶标。免疫印迹法证实了靶点之一的核酮糖-1,5-二磷酸羧化酶/加氧酶（Rubisco）的体内和体外亚硝基化作用。Rubisco在光合作用中起着核心作用，利用(NaHCO3)-C-14测定了s -亚硝基化对其酶活性的影响。释放no的化合物s -亚硝基谷胱甘肽以剂量依赖的方式抑制其活性，表明首次通过亚硝基化灭活Rubisco。
Nitric oxide (NO) is a signaling molecule that affects a myriad of processes in plants. However, the mechanistic details are limited. NO post-translationally modifies proteins by S-nitrosylation of cysteines. The soluble S-nitrosoproteome of a medicinal, crassulacean acid metabolism (CAM) plant, Kalanchoe pinnata, was purified using the biotin switch technique. Nineteen targets were identified by MALDI-TOF mass spectrometry, including proteins associated with carbon, nitrogen and sulfur metabolism, the cytoskeleton, stress and photosynthesis. Some were similar to those previously identified in Arabidopsis thaliana, but kinesin-like protein, glycolate oxidase, putative UDP glucose 4-epimerase and putative DNA topoisomerase II had not been identified as targets previously for any organism. In vitro and in vivo nitrosylation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), one of the targets, was confirmed by immunoblotting. Rubisco plays a central role in photosynthesis, and the effect of S-nitrosylation on its enzymatic activity was determined using (NaHCO3)-C-14. The NO-releasing compound S-nitrosoglutathione inhibited its activity in a dose-dependent manner suggesting Rubisco inactivation by nitrosylation for the first time.