Cold stress changes the concanavalin A-positive glycosylation pattern of proteins expressed in the basal parts of rice leaf sheaths
Cold stress changes the concanavalin A-positive glycosylation pattern of proteins expressed in the basal parts of rice leaf sheaths
复制标题
DOI：
10.1007/s00726-008-0039-4
复制
发表时间：
2009-01-01
期刊：
影响因子：
3.5
通讯作者：
Furukawa, K.
中科院分区：
文献类型：
--
作者：
Komatsu, S.;Yamada, E.;Furukawa, K.
翻译后修饰如糖基化对于改变蛋白质的性质和功能是重要的。为了分析糖基化在水稻冷胁迫过程中的重要性，采用蛋白质组学方法。从水稻叶鞘基部提取的蛋白质经双向聚丙烯酰胺凝胶电泳分离后，用刀豆球蛋白A进行凝集素印迹分析。从总共检测到的250个蛋白中，有22个与凝集素反应，表明它们是N-糖基化蛋白。为了确定这些蛋白质的N-糖基化如何受到冷胁迫的影响，水稻幼苗在5A摄氏度下孵育48小时，从叶鞘基部提取的蛋白质通过凝集素印迹分析。冷应激改变了22种糖蛋白中的12种对凝集素的反应性。通过蛋白质测序和质谱法确定了12种蛋白质的身份，其中大多数糖蛋白被归类为参与能量产生。此外，钙网蛋白，12种糖蛋白之一，也被磷酸化作为冷胁迫的结果。这些结果表明，水稻叶鞘基部的冷胁迫改变了钙网蛋白的糖基化和磷酸化谱，钙网蛋白是调节其他蛋白质质量控制的关键蛋白质。
Post-translational modifications such as glycosylation are important for changing the properties and functions of proteins. To analyze the importance of glycosylation during cold stress in rice, a proteomics approach was used. Proteins extracted from the basal part of rice leaf sheaths were separated by two-dimensional polyacrylamide gel electrophoresis, and subjected to lectin blot analysis using concanavalin A. From a total of 250 detected proteins, 22 reacted with the lectin, suggesting that they were N-glycosylated proteins. To determine how N-glycosylation of these proteins is affected by cold stress, rice seedlings were incubated at 5A degrees C for 48 h, and proteins extracted from the basal parts of leaf sheaths were analyzed by the lectin blot assay. Cold stress changed the reactivity toward the lectin for 12 of the 22 glycoproteins. The identity of the 12 proteins was determined by protein sequencing and mass spectrometry with the majority of these glycoproteins being categorized as involved in energy production. Furthermore, calreticulin, one of the 12 glycoproteins, was also phosphorylated as a result of cold stress. These results indicate that cold stress of the basal parts of rice leaf sheaths changes the glycosylation and phosphorylation profiles of calreticulin, a key protein that regulates the quality control of other proteins.