An in vivo chemical genetic approach for targeted glycoproteome analysis in Drosophila melanogaster
Schmidt, S. D.; Alexandre, C.; Di Vagno, L.; Zhang, L.; Flynn, H.; Kurth, J.; Murray, S.; Ruiz Herrera, S.; Shaw, H.; Bineva-Todd, G.; Yilmaz Tastan, O.; Skehel, M.; Samara, N. L.; Ten Hagen, K.; Vincent, J.-P.; Schumann, B.
Show abstract
Glycans impact every aspect of physiology. Despite the rapid increase in relevance and translational potential, there remains a lack of tools for studying glycosylation in vivo. Here, we present FlyMOE, a chemical-genetic platform for targeted glycoprotein profiling in Drosophila melanogaster. Transgenic flies are equipped with biosynthetic capabilities to install bioorthogonally-tagged monosaccharides into the cellular glycoproteome. The genetic tractability of Drosophila allows for versatile application based on developmental stages, genetic drivers and glycosyltransferase enzymes. We employ FlyMOE to create an overview of the glycoproteome of Drosophila tissues and embryos based on mass spectrometry glycoproteomics, and investigate the substrates of individual members of the PGANT glycosyltransferase family in vivo. FlyMOE will allow the detailed investigation of the glycoproteome in Drosophila and similarly genetically modifiable in vivo model systems.
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