Two-component high-affinity nitrate transport system in barley: Membrane localization, protein expression in roots and a direct protein-protein interaction
Two-component high-affinity nitrate transport system in barley: Membrane localization, protein expression in roots and a direct protein-protein interaction
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DOI：
10.5511/plantbiotechnology.26.197
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发表时间：
2009-03-25
影响因子：
1.6
通讯作者：
Sueyoshi, Kuni
中科院分区：
文献类型：
--
作者：
Ishikawa, Shinji;Ito, Yuka;Sueyoshi, Kuni
植物中硝酸盐的高亲和力运输系统（HATS）由双组分NRT 2/NAR 2运输系统操作。然而，NRT 2和NAR 2在蛋白质水平上的调控和定位在很大程度上是未知的，尤其是在作物物种中。在这项研究中，大麦（大麦），膜定位，蛋白质表达在根中，和直接的蛋白质-蛋白质相互作用的HvNRT 2和HvNAR 2蛋白进行了研究。免疫化学分析表明，HvNRT 2和HvNAR 2蛋白共定位于质膜的根。HvNRT 2和HvNAR 2蛋白的表达更强烈地诱导处理与较高浓度的外部硝酸盐，而HATS活性和转录HvNRT 2和HvNAR 2显着抑制。使用重组蛋白的亲和柱结合分析表明，HvNRT 2.1的C-末端可能参与其与HvNAR 2.3中心区域的结合，并且存在于HvNRT 2.1 C-末端中的Ser 463在结合能力中起作用。
A high affinity transport system (HATS) for nitrate in plants is operated by a two-component NRT2/NAR2 transport system. However, the regulation and localization of NRT2 and NAR2 at protein level are largely unknown and especially so in crop plant species. In this study with barley (Hordeum vulgare), membrane localization, protein expression in the roots, and a direct protein-protein interaction of HvNRT2 and HvNAR2 proteins were investigated. Immunochemical analysis showed that both HvNRT2 and HvNAR2 proteins were co-localized in the plasma membrane of the roots. Expression of HvNRT2 and HvNAR2 proteins was more strongly induced by treatment with higher concentrations of external nitrate, while HATS activity and transcripts for HvNRT2 and HvNAR2 were markedly repressed. An affinity Column binding analysis using recombinant proteins suggests that the C-terminus of HvNRT2.1 is possibly involved in its binding to the HvNAR2.3 central region and that the Ser463 present in the HvNRT2.1 C-terminus plays a role in the binding ability.