The role of tonoplast protein phosphorylation in activity regulation and protein targeting

液泡膜蛋白磷酸化在活性调节和蛋白靶向中的作用

• 批准号: 214983253
• 负责人: Professorin Dr. Waltraud Schulze
• 金额: --
• 依托单位: Fachgebiet Systembiologie der Pflanze (190d)
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/214983253?language=en
• 关键词: role; tonoplast; protein; phosphorylation; activity

The tonoplast membrane is an important compartment across which solute transport is tightly controlled under various environmental conditions. The goal of the proposed experiments is to elucidate the role of tonoplast protein phosphorylation in regulation of vacuolar solute accumulation and in protein targeting by the following experiments: (i) A targeted analysis by single reaction monitoring using specific labelled standard peptides of already known and predicted phosphopeptides and respective non-phosphopeptides will allow us to compare phosphorylation status and protein abundance. Up to 20 key transporters and channels across a wide range of stress conditions will be analyzed. (ii) An untargeted “discovery” approach using 15N-labeing will be carried out to identify proteins for which specific stress conditions or mutations of protein targeting pathways result in altered subcellular membrane location. Finally, different phosphorylation sites of the TMT protein family will be used as an example for (iii) in-depth characterization of the role of protein phosphorylation in stress-induced solute accumulation and protein targeting.

液泡膜是一个重要的隔室，在各种环境条件下，溶质的运输在其上受到严格控制。拟议实验的目标是通过以下实验阐明液泡膜蛋白磷酸化在调节液泡溶质积累和蛋白质靶向中的作用：（i）使用已知和预测的磷酸肽和相应非磷酸肽的特定标记标准肽通过单反应监测进行靶向分析将使我们能够比较磷酸化状态和蛋白质丰度。将分析各种压力条件下的多达 20 个关键转运蛋白和通道。 (ii) 将采用 15N 标记的非靶向“发现”方法来识别特定应激条件或蛋白质靶向途径突变导致亚细胞膜位置改变的蛋白质。最后，以TMT蛋白家族的不同磷酸化位点为例，(iii)深入表征蛋白磷酸化在应激诱导的溶质积累和蛋白靶向中的作用。

项目成果

Databases for plant phosphoproteomics.

• DOI: 10.1007/978-1-4939-2648-0_16
• 发表时间: 2015
• 期刊: Methods in molecular biology
• 作者: W. Schulze;Q. Yao;Dong Xu

From Phosphoproteome to Modeling of Plant Signaling Pathways.

从磷酸化蛋白质组到植物信号通路建模

• DOI: 10.1007/978-1-4939-3341-9_18
• 发表时间: 2016
• 期刊: Methods in molecular biology
• 作者: Zakhartsev M;Pertl-Obermeyer H;Schulze WX
• 通讯作者: Schulze WX

In vivo cross-linking combined with mass spectrometry analysis reveals receptor-like kinases and Ca(2+) signalling proteins as putative interaction partners of pollen plasma membrane H(+) ATPases.

体内交联与质谱分析相结合揭示了受体样激酶和 Ca(2 ) 信号蛋白作为花粉质膜 H( ) ATP 酶的假定相互作用伙伴

• DOI: 10.1016/j.jprot.2014.05.001
• 发表时间: 2014
• 期刊: Journal of proteomics
• 影响因子: 3.3
• 作者: Pertl-Obermeyer;Schulze;Obermeyer
• 通讯作者: Obermeyer

Kinase activity and specificity assay using synthetic peptides.

使用合成肽进行激酶活性和特异性测定

• DOI: 10.1007/978-1-4939-2648-0_7
• 发表时间: 2015
• 期刊: Methods in molecular biology
• 作者: Schulze