Identification of tonoplast membrane proteins involved in cellular response to environmental stress by quantitative/comparative- and phospho-proteomics

通过定量/比较和磷酸蛋白质组学鉴定参与细胞对环境应激反应的液泡膜蛋白

• 批准号: 76388309
• 负责人: Dr. Oliver Trentmann
• 金额: --
• 依托单位: Fachgebiet Pflanzenphysiologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/76388309?language=en
• 关键词: Identification; tonoplast; membrane; proteins; involved

Proteom analyses represent a formidable tool to identify vacuolar proteins and to enlighten their relevance in stress response. During the past period of the research grant we have mainly focused on establishing a platform to isolate highly pure vacuoles applicable for mass spectrometry based proteomics. We isolated vacuoles from cold incubated and control plants to investigate changes in the tonoplast proteome during the process of cold acclimation. Thereby several changes in the abundance of tonoplast proteins and alterations in protein phosphorylation were identified providing first insights into the physiological relevance of the vacuole on cold acclimation. By these proteome analyses we identified several unknown proteins residing in the tonoplast and characterized one of these in detail. The tonoplast copper transporter COPT5 belongs to the eukaryotic Ctr family, catalyses the export of copper ions from the vacuole and is responsible for the inter organ allocation of copper in Arabidopsis. In the proposed second funding period we will expand the comparative analysis of changes in the proteome of the tonoplast induced by other abiotic factors like drought, salt, heat and osmotic stress. In parallel we will investigate how such stress factors alter the phosphorylation of tonoplast proteins to understand the relevance of this post-translational protein modification on the regulation of tonoplast transport mechanisms. Finally, we will investigate the relevance of individual tonoplast proteins for the plants stress response and we will effort to clarify the biochemical and physiological function of so far unknown tonoplast proteins.

蛋白质分析代表了识别液泡蛋白并启发其在压力反应中的相关性的强大工具。在研究赠款的过去期间，我们主要集中于建立一个平台，以隔离适用于质谱蛋白质组学的高度纯净液泡。我们从冷孵化和控制植物中分离出液泡，以研究冷适应过程中块状蛋白质组的变化。从而确定了块状蛋白质的丰度和蛋白质磷酸化的改变，从而提供了对液泡在冷适应中的生理相关性的首先见解。通过这些蛋白质组分析，我们鉴定了几种位于块状体中的未知蛋白质，并详细介绍了其中一种。托糖铜转运蛋白COPT5属于真核CTR家族，催化了液泡的铜离子出口，并负责拟南芥中铜的间分配。在拟议的第二融资期间，我们将扩大其他非生物因素（如干旱，盐，热和渗透压）引起的块状蛋白质组变化的比较分析。同时，我们将研究这种应力因素如何改变块状蛋白质的磷酸化，以了解这种翻译后蛋白质修饰与调节托管传输机制的相关性。最后，我们将研究单个块状蛋白与植物压力反应的相关性，我们将努力阐明迄今未知骨体蛋白的生化和生理功能。