Genome-wide activity-based profiling of Arabidopsis proteomes

• Antragsteller: Professor Dr. Renier A.L. van der Hoorn; Professor Dr. Markus Kaiser; Dr. Jürgen Schmidt
• Fachliche Zuordnung: Zell- und Entwicklungsbiologie der Pflanzen
• Förderung: Förderung von 2007 bis 2011
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 50069751

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.

• DFG-Verfahren: Sachbeihilfen