Proteomic analysis of the effect of heat stress on hexaploid wheat grain:: Characterization of heat-responsive proteins from non-prolamins fraction
Proteomic analysis of the effect of heat stress on hexaploid wheat grain:: Characterization of heat-responsive proteins from non-prolamins fraction
复制标题
DOI：
10.1002/pmic.200300570
复制
发表时间：
2004-02-01
期刊：
影响因子：
3.4
通讯作者：
Branlard, G
中科院分区：
文献类型：
--
作者：
Majoul, T;Bancel, E;Branlard, G
热胁迫对小麦六倍体籽粒蛋白质组的影响是我们研究的热点。蛋白质组学工具可以表征总胚乳的热响应蛋白，主要由蛋白组成。本文的工作完成了这一研究；我们的目的是分析热应激对主要由白蛋白和球蛋白组成的水溶性部分的影响。这些蛋白通过双向电泳（2-DE）分离，考马斯亮蓝（CBB）染色显示，Melanie-3软件分析。43个热变蛋白中，有24个蛋白表达上调，19个蛋白表达下调。利用基质辅助激光解吸/电离飞行时间质谱法（MALDI-TOF-MS）对这些蛋白进行了数据库检索，鉴定出42个位点。其中一些是参与植物不同代谢途径的酶，如参与淀粉合成途径的颗粒结合淀粉合成酶和葡萄糖-1-磷酸腺苷转移酶；与碳水化合物代谢有关的-淀粉酶，以及与四种减热蛋白有关的ATP合成酶-链。此外，5个热上调蛋白与小热休克蛋白相似，另外3个位点与延伸因子或真核生物翻译起始因子有关。还鉴定并讨论了参与非生物胁迫或植物防御机制的蛋白质。
The effect of heat stress on hexaploid wheat grain proteome was recently analyzed in our previous works. Proteomic tools allowed the characterization of heat-responsive proteins of total endosperm, composed mainly of prolamins. The present work completes this study; our aim was to analyze the effect of heat stress on the water-soluble fraction, composed essentially of albumins and globulins. These proteins were separated by two-dimensional electrophoresis (2-DE), visualized by Coomassie Brilliant Blue (CBB) staining and analyzed by Melanie-3 software. Of the 43 heat-changed proteins, 24 were found to be up-regulated whereas 19 spot proteins were down-regulated. All of these proteins were subjected to matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) followed by database searching which allowed the identification of 42 spots. Of these, some were enzymes involved in different metabolic pathways of plants, such as granule-bound starch synthase and glucose-1-phosphate adenyltransferase, involved in the starch synthesis pathway; beta-amylase, involved in carbohydrate metabolism, and the ATP synthase beta-chain that was related to four heat-decreased proteins. Moreover, five heat up-regulated proteins showed similarities with small heat shock proteins while three other spots were related to elongation factors or eucaryotic translation initiation factors. Proteins involved in abiotic stresses or in plant defense mechanism were also identified and are discussed.