iTRAQ-based quantitative proteome and phosphoprotein characterization reveals the central metabolism changes involved in wheat grain development.
iTRAQ-based quantitative proteome and phosphoprotein characterization reveals the central metabolism changes involved in wheat grain development.
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基于 iTRAQ 的定量蛋白质组和磷蛋白表征揭示了小麦籽粒发育中涉及的中心代谢变化
DOI：
10.1186/1471-2164-15-1029
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发表时间：
2014-11-27
期刊：
影响因子：
4.4
通讯作者：
Yan Y
中科院分区：
文献类型：
--
作者：
Ma C;Zhou J;Chen G;Bian Y;Lv D;Li X;Wang Z;Yan Y
小麦（Triticum aestivum L.）是重要的经济作物。基于二维凝胶的方法受到蛋白质鉴定率低和缺乏准确蛋白质定量的限制。最近开发的同量异位素标签相对和绝对定量（iTRAQ）方法可以灵敏和准确的蛋白质定量。在这里，我们进行了第一个基于iTRAQ的定量蛋白质组和磷酸化蛋白质分析在小麦籽粒发育。采用iTRAQ定量蛋白质组技术、TiO 2微柱技术和液相色谱-串联质谱（LC-MS/MS）技术，对中国面包小麦优良品种盐优361籽粒发育过程中代谢蛋白的蛋白质组谱和磷蛋白特征进行了研究。在鉴定的1,146个非冗余蛋白中，421个在丰度上显示出至少2倍的差异，并且它们被鉴定为差异表达蛋白（DEP），包括256个上调蛋白和165个下调蛋白。在421个DEPs中，鉴定出6种蛋白表达模式，其中大部分为上调、下调和上调-下调表达模式。421个DEPs主要分布于细胞膜和细胞质中，参与不同的代谢过程。系统聚类分析表明，与淀粉合成、贮藏蛋白和防御/胁迫相关蛋白相关的DEPs在籽粒发育后期显著积累，而与蛋白质合成/组装/降解和光合作用相关的DEPs在籽粒发育过程中表现出相反的表达模式。对编码不同代谢蛋白的12个代表性基因的实时定量聚合酶链反应（qRT-PCR）分析表明，在籽粒发育过程中存在一定的转录和翻译表达差异。磷酸化蛋白质分析表明，23个DEPs，如AGEs，蔗糖合成酶，热休克蛋白90，丝氨酸蛋白酶抑制剂磷酸化在发育中的籽粒，主要参与淀粉合成和胁迫/防御。我们的研究结果揭示了小麦籽粒发育过程中复杂的定量蛋白质组和磷酸化谱。许多DEPs参与小麦籽粒淀粉和蛋白质的合成以及逆境防御，是小麦产量和品质的重要基础。特别是一些参与淀粉合成和胁迫/防御的关键DEPs被磷酸化，这表明它们在小麦籽粒发育中的作用。本文的在线版本（doi：10.1186/1471-2164-15-1029）包含补充材料，可供授权用户使用。
Wheat (Triticum aestivum L.) is an economically important grain crop. Two-dimensional gel-based approaches are limited by the low identification rate of proteins and lack of accurate protein quantitation. The recently developed isobaric tag for relative and absolute quantitation (iTRAQ) method allows sensitive and accurate protein quantification. Here, we performed the first iTRAQ-based quantitative proteome and phosphorylated proteins analyses during wheat grain development. The proteome profiles and phosphoprotein characterization of the metabolic proteins during grain development of the elite Chinese bread wheat cultivar Yanyou 361 were studied using the iTRAQ-based quantitative proteome approach, TiO2 microcolumns, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Among 1,146 non-redundant proteins identified, 421 showed at least 2-fold differences in abundance, and they were identified as differentially expressed proteins (DEPs), including 256 upregulated and 165 downregulated proteins. Of the 421 DEPs, six protein expression patterns were identified, most of which were up, down, and up-down expression patterns. The 421 DEPs were classified into nine functional categories mainly involved in different metabolic processes and located in the membrane and cytoplasm. Hierarchical clustering analysis indicated that the DEPs involved in starch biosynthesis, storage proteins, and defense/stress-related proteins significantly accumulated at the late grain development stages, while those related to protein synthesis/assembly/degradation and photosynthesis showed an opposite expression model during grain development. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of 12 representative genes encoding different metabolic proteins showed certain transcriptional and translational expression differences during grain development. Phosphorylated proteins analyses demonstrated that 23 DEPs such as AGPase, sucrose synthase, Hsp90, and serpins were phosphorylated in the developing grains and were mainly involved in starch biosynthesis and stress/defense. Our results revealed a complex quantitative proteome and phosphorylation profile during wheat grain development. Numerous DEPs are involved in grain starch and protein syntheses as well as adverse defense, which set an important basis for wheat yield and quality. Particularly, some key DEPs involved in starch biosynthesis and stress/defense were phosphorylated, suggesting their roles in wheat grain development. The online version of this article (doi:10.1186/1471-2164-15-1029) contains supplementary material, which is available to authorized users.
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中国小麦优良品种小偃6号籽粒发育过程中白蛋白和球蛋白动态
影响因子：
3.8
作者：
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影响因子：
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DOI：
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发表时间：
1984-01-01
期刊：
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影响因子：
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作者：
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通讯作者：
CUMING, AC
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影响因子：
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作者：
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