Identification and Absolute Quantitation of Protein Phosphorylation Networks in Oilseeds

油籽中蛋白质磷酸化网络的鉴定和绝对定量研究[j]: Jay J. Thelen，密苏里大学哥伦比亚分校生物化学系合作者：徐东，密苏里大学哥伦比亚分校计算机科学系]植物种子是重要的可再生天然产物，如油、蛋白质、淀粉和纤维。虽然这些储存化合物的生物合成途径是已知的，但不清楚这些途径是如何在油籽中被调节的，油籽产生更多的油和蛋白质。蛋白质磷酸化是许多代谢酶的重要调控机制，初步数据显示油菜和大豆发育过程中存在数百种磷酸化蛋白。对三种不同油籽发育过程中磷蛋白调控机制的鉴定和量化，将为了解油籽发育过程中的调控网络提供基础，并有助于制定新的作物改良策略。本研究旨在揭示三种不同油籽植物（油菜、大豆和拟南芥）在种子发育的油脂和蛋白质积累阶段主要参与代谢过程调节的磷酸化蛋白和蛋白激酶网络。除了这个基本的科学目标，该提案的更广泛的影响包括：(1)组织以蛋白质组学为主题的研讨会和开发基于网络的蛋白质组学教程，分别针对高中教师和学生；(2)培养蛋白质组学这一新兴学科的本科至博士后水平的植物科学家；(3)通过大学资助的本科生研究项目和奖学金招收代表性不足的学生；(4)利用新兴的绝对定量（AQUA）肽技术开发磷酸肽鉴定和定量技术；(5)为植物蛋白磷酸化群落生成合成AQUA肽工具；(6)以多种用户直观的形式公开传播磷蛋白质组学数据，为科学界修改种子代谢以造福健康和环境提供新的方向。有关网站请参阅http://oilseedproteomics.missouri.edu

IDENTIFICATION AND ABSOLUTE QUANTITATION OF PROTEIN PHOSPHORYLATION NETWORKS IN OILSEEDSPI: Jay J. Thelen, Department of Biochemistry, University of Missouri-ColumbiaCollaborator: Dong Xu, Computer Science Department, University of Missouri ColumbiaPlant seeds are important, renewable sources of natural products such as oil, protein, starch and fiber. Though the biosynthetic pathways for these storage compounds are mostly known, it is not clear how these pathways are regulated in oilseeds which produce higher quantities of oil and protein. Protein phosphorylation is an important regulatory mechanism for many metabolic enzymes and preliminary data reveal hundreds of phosphoproteins in developing oilseed rape and soybean. Identification and quantification of phosphoprotein control mechanisms in developing seed of three different oilseeds will provide the basis for understanding the regulatory networks involved in oilseed development and will help in designing new strategies for crop improvement. This proposal aims to unravel networks of phosphoproteins and protein kinases primarily involved in the regulation of metabolic processes during the oil and protein accumulation phase of seed development in three different oilseed plants, oilseed rape, soybean, and Arabidopsis thaliana. In addition to this fundamental scientific objective, the broader impacts of this proposal include: (1) organizing proteomics-themed workshops and developing web-based proteomics tutorials targeting High School teachers and students, respectively; (2) training plant scientists at the undergraduate through post-doctoral levels in the emerging discipline of proteomics; (3) recruiting under-represented students through University-sponsored undergraduate research programs and fellowships; (4) developing techniques for phosphopeptide identification and quantification using the emerging technology of absolute quantification (AQUA) peptides; (5) generating synthetic AQUA peptide tools for the plant protein phosphorylation community; and (6) public dissemination of phosphoproteomics data in multiple, user-intuitive formats to provide new directions for the scientific community to modify seed metabolism for the benefit of health and environment. For website see http://oilseedproteomics.missouri.edu