Comprehensive phosphoproteome analysis in rice and identification of phosphoproteins responsive to different hormones/stresses
Comprehensive phosphoproteome analysis in rice and identification of phosphoproteins responsive to different hormones/stresses
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DOI：
10.1021/pr0501160
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发表时间：
2005-09-01
影响因子：
4.4
通讯作者：
Komatsu, S
中科院分区：
文献类型：
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作者：
Khan, M;Takasaki, H;Komatsu, S
采用体外蛋白磷酸化和双向凝胶电泳法对大米中的磷蛋白进行检测。水稻叶鞘粗提物体外蛋白质磷酸化后，在2D-Gel上检测到44个磷蛋白。通过Q-TOF MS/MS和/或MALDI-TOF MS分析，鉴定出42个磷蛋白，其中与信号转导有关的比例最大(30%)，参与代谢的比例最大(18%)。用不同激素和胁迫处理水稻幼苗时，观察到不同激素和胁迫处理对13种蛋白质的磷酸化有不同程度的增强作用。此外，当比较水稻叶鞘、叶片和根中激素/胁迫调节的磷蛋白时，发现所有组织中只有细胞质苹果酸脱氢酶被磷酸化。结果表明，在水稻的磷酸化级联反应中，糖酵解代谢过程和钙信号似乎是响应激素和逆境的重要靶标。此外，通过P-32标记直接显示磷蛋白并进行质谱鉴定，为分析水稻磷蛋白质组提供了一种准确可靠的方法。
Phosphoproteins in rice were detected by in vitro protein phosphorylation followed by two-dimensional polyacrylamide gel electrophoresis. Forty-four phosphoproteins were detected on a 2D-gel after in vitro protein phosphorylation of the crude extract from rice leaf sheath. Among the phosphoproteins detected, 42 were identified through analysis by Q-TOF MS/MS and/or MALDI-TOF MS. The largest percentage of the identified phosphoproteins are involved in signaling (30%), while 18% are involved in metabolism. When rice seedlings were treated with various hormones and stresses, it was observed that the phosphorylation of 13 proteins was enhanced differentially by different hormone and stress treatments. Furthermore, when the hormone/stress regulated phosphoproteins are compared in rice leaf sheath, leaf blade and root, only cytoplasmic malate dehydrogenase was found to be phosphorylated in all the tissues. Results suggest that in the phosphorylation cascade of rice, glycolytic metabolism processes and Ca2+-Signaling seem to be important targets in response to hormones and stresses. Furthermore, the direct visualization of phosphoproteins by P-32-labeling and their mass spectrometric identification provides an accurate and reliable method of analyzing the rice phosphoproteome.