Chapter Fourteen - The Glycomics of Glycan Glucuronylation in Drosophila melanogaster

Abstract

As glycan characterization methods increase in sensitivity, new opportunities arise to undertake glycomic analyses on limiting amounts of material. Developing systems present special challenges since the amount of available tissue can restrict deep glycan characterization. We have optimized mass spectrometric methods with the goal of obtaining full glycan profiles from small amounts of tissue derived from organisms of particular interest. A major target of our efforts has been the Drosophila embryo, allowing us to leverage the tools already developed in this organism to meld glycomics, genomics, and molecular genetics. Our analysis of the N-linked, O-linked (non-GAG), and glycosphingolipid (GSL) glycans of the Drosophila embryo have identified expected and unexpected glycan structures. We have verified previous findings regarding the predominance of high-Man and pauci-Man N-linked glycans, but have also detected minor families of sialylated and glucuronylated N-linked structures. Glucuronic acid (GlcA) also presents itself as an abundant modification of O-linked and GSL glycans. We describe critical advancements in our methodology and present the broad range of contexts in which GlcA is found in the Drosophila embryo.