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.

蛋白质组分析代表了一个强大的工具，以确定液泡蛋白，并启发他们在应激反应的相关性。在过去的研究资助期间，我们主要集中在建立一个平台，以分离适用于基于质谱的蛋白质组学的高纯液泡。我们从冷培养和对照植物中分离液泡，以研究冷驯化过程中液泡膜蛋白质组的变化。因此，确定了液泡膜蛋白丰度的几种变化和蛋白磷酸化的改变，首次深入了解了液泡对冷驯化的生理相关性。通过这些蛋白质组分析，我们确定了几个未知的蛋白质居住在液泡膜，并详细描述了其中之一。液泡膜铜转运蛋白COPT 5属于真核细胞Ctr家族，催化铜离子从液泡中输出，并负责拟南芥中铜的器官间分配。在拟议的第二个资助期内，我们将扩大由其他非生物因素（如干旱，盐，热和渗透胁迫）引起的液泡膜蛋白质组变化的比较分析。与此同时，我们将研究这些应激因素如何改变液泡膜蛋白的磷酸化，以了解这种翻译后蛋白修饰对液泡膜转运机制的调节的相关性。最后，我们将研究单个液泡膜蛋白与植物胁迫反应的相关性，并努力阐明迄今未知的液泡膜蛋白的生化和生理功能。