Genome-wide activity-based profiling of Arabidopsis proteomes

基于全基因组活性的拟南芥蛋白质组分析

• 批准号: 50069751
• 负责人: Professor Dr. Renier A.L. van der Hoorn
• 金额: --
• 依托单位: Max-Planck-Institut für Pflanzenzüchtungsforschung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/50069751?language=en
• 关键词: Genome; wide; activity; based; profiling

Information on protein activity is essential to the annotation and understanding of protein functions since it is the activity of a protein, rather than its abundance, that ultimately dictates its role in a living organism. We aim to uncover the proteome activity layer of Arabidopsis through a novel, genome-wide technology called activity-based proteome profiling (ABPP). ABPP can be applied on living tissues or on protein extracts and is based on labeled small molecules (probes) that react with active site residues of a broad range of proteins in an activity-dependent manner. Labeled proteins can be separated and visualized on protein gels and identified by mass spectrometry (MS). This project aims to generate activity information of 100-1000 different proteins from various mechanistic classes from different Arabidopsis proteomes through an intimate interdisciplinary collaboration between an organic chemist, biochemist and MS-specialist. Activities will be characterized and confirmed by heterologous expression. Labeling sites of as yet non-annotated proteins will be identified and validated to reveal their catalytic mechanisms. Finally, inhibitory signatures will be generated to sub-classify a large redundant protein family into functional units. Data will be shared through an open-access searchable database where plant scientists can retrieve activity information for their favorite protein(s) and request probes and protocols for further studies.

关于蛋白质活性的信息对于蛋白质功能的注释和理解是必不可少的，因为它是蛋白质的活性，而不是它的丰度，最终决定了它在生物体中的作用。我们的目标是揭示蛋白质组活性层的拟南芥通过一种新的，全基因组技术称为基于活性的蛋白质组分析（ABPP）。ABPP可应用于活组织或蛋白质提取物，并基于标记的小分子（探针），其以活性依赖性方式与广泛蛋白质的活性位点残基反应。标记的蛋白质可以在蛋白质凝胶上分离和可视化，并通过质谱法（MS）鉴定。该项目旨在通过有机化学家，生物化学家和MS专家之间的密切跨学科合作，从不同的拟南芥蛋白质组中产生100-1000种不同蛋白质的活性信息。活性将通过异源表达进行表征和确认。尚未注释的蛋白质的标记位点将被识别和验证，以揭示其催化机制。最后，将生成抑制签名以将大的冗余蛋白质家族细分为功能单元。数据将通过一个开放访问的可搜索数据库共享，植物科学家可以检索他们最喜欢的蛋白质的活性信息，并请求探针和协议进行进一步研究。