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Glycosylation of peptide hormones and extensins fr.. (ExHoMo)
Glycosylation of peptide hormones and extensins from the model plant Arabidopsis thaliana and in-vitro synthesis of glycosylated peptides
(ExHoMo)
Date du début: 1 mai 2012,
Date de fin: 30 sept. 2014
PROJET
TERMINÉ
As global food prices mount, the imperative for novel approaches to crop growth becomes evident. A recently discovered group of glycosylated peptides with hormonal characteristics has a similar impact on plant growth and environmental response as the classical phytohormones that were pivotal in the first green revolution.We aim to develop methods for the in-vitro synthesis of these hormones.Glycosylation is essential for peptide function. Identification of glycosyl transferases (GTs) acting on these peptides is a prerequisite for in-vitro synthesis. I will identify potential GTs by combining existing bioinformatic techniques and genomic data available at the host institution with proteomic data that I bring with me from my previous post-doctoral position in the US. I will also search for the last unidentified GT initiating glycosylation in a related, non-hormonal group of secreted peptides.Working with molecular biologists at the host institution, I will develop my molecular skills constructing tagged synthetic peptides and expressing these in putative GT mutants selected from the proteomic and genomic data. Working with mass spectrometry specialists I will analyse glycosylation signatures on peptides retrieved from wild type and mutant plants.Simultaneously, the fate of synthetic peptides progressing through the secretory system will be followed using protocols I developed during my time in the US and equipment that I bring with me. This will establish the distribution of GTs throughout the secretory system and act as a proof of concept for this technique in analysing modifications on other secreted proteins.Putative GTs expressed in heterologous systems with specific donor and acceptor substrates will confirm GT identity and demonstrate in-vitro synthetic capacity, a move towards developing novel agricultural tools.This project will develop state-of-the-art methods with multiple applications and mark a EU contribution to an exciting new field.
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