Proteomics approach to identify wound‐response related proteins from rice leaf sheath
Proteomics approach to identify wound‐response related proteins from rice leaf sheath
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DOI：
10.1002/pmic.200390066
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发表时间：
2003-04
期刊：
影响因子：
3.4
通讯作者：
S. Shen;Y. Jing;T. Kuang
中科院分区：
文献类型：
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作者：
S. Shen;Y. Jing;T. Kuang
为了避免完整植物的复杂条件，以简单地分析蛋白质在创伤反应胁迫下，我们使用了离体水稻叶鞘，这是水稻幼苗的一个非常活跃的部分。以水稻叶鞘为材料，分别在切割后0、12、24、48 h提取蛋白质，进行双向聚丙烯酰胺凝胶电泳。差异显示的蛋白质的变化被发现在叶鞘切割后的0-48 h的时间进程。与4种二维凝胶相比，10种蛋白质上调，19种蛋白质下调。其中，14个蛋白质通过N末端或内部氨基酸序列进行了分析。其中9个蛋白质的功能比较明确。六种蛋白质由于其封闭的N末端而没有产生氨基酸序列信息。此外，通过基质辅助激光解吸/电离-飞行时间质谱法测定了11种蛋白质，并鉴定了蛋白质数据库匹配。结果表明，下调的蛋白质是钙网蛋白（nos. 5、6）、组蛋白H1（编号15）和血红蛋白（编号17）、推定的过氧化物酶（编号19）;上调的蛋白质为Bowman-比尔克胰蛋白酶抑制剂（编号23）、推定的受体样蛋白激酶（编号17）、推定的过氧化物酶（编号19）。24，25），钙调蛋白相关蛋白（编号26），核酮糖-1，5-二磷酸羧化酶/加氧酶的小亚基（编号27），甘露糖结合水稻凝集素（编号27），28、29）。在所有上述蛋白质中，四个（nos. 23，24，25，26）已被证实是创伤反应蛋白。不能排除其他也与伤口反应相关的蛋白质，如信号转导相关蛋白质（nos. 5，6），光合作用相关蛋白（编号27），和应激反应蛋白（编号27）。19、28、29）。这是第一次在水稻叶鞘蛋白质的变化响应创伤已被证明。
In order to avoid the complex conditions of the intact plant for simple analysis of proteins in wound‐response stress, we used the detached rice leaf sheath which is a very active part of the rice seedling. Proteins were extracted from rice leaf sheath at 0, 12, 24, 48 h after cutting and separated by two‐dimensional (2‐D) polyacrylamide gel electrophoresis. Changes in differentially displayed proteins were found in leaf sheaths after cutting in the 0–48 h time course. Ten proteins were up‐regulated, while 19 proteins were down‐regulated compared with those on the four 2‐D gels. Among them, 14 proteins were analyzed by N‐terminal, or internal amino acid sequence. The clear functions of nine proteins could be identified. Six proteins did not yield amino acid sequence information due to their blocked N‐termini. Furthermore, 11 proteins were determined by matrix‐assisted laser desorption/ionization‐time of flight mass spectrometry, and identified protein database matching. It was shown that the down‐regulated proteins were calreticulin (nos. 5, 6), histone H1 (no. 15) and hemoglobin (no. 17), putative peroxidase (no. 19); the up‐regulated proteins were Bowman‐Birk trypsin inhibitor (no. 23), putative receptor‐like protein kinase (nos. 24, 25), calmodulin‐related protein (no. 26), small subunit of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (no. 27), mannose‐binding rice lectin (nos. 28, 29). Among all the above proteins, four (nos. 23, 24, 25, 26) have been confirmed to be wound‐response proteins. The others cannot be excluded as also being related to wound‐responses, such as the signal transduction‐related proteins (nos. 5, 6), photosynthesis‐related protein (no. 27), and stress‐response proteins (nos. 19, 28, 29). This is the first time protein changes in response to wounding in rice leaf sheath have been shown.