Investigating Glycosylation and Phosphorylcholine Post-Translational Modifications in the Free-Living Nematode, Caenorhabditis elegans
AuthorSnodgrass, Casey J.
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<italic>Caenorhabditis elegans</italic> is a classic genetic model, but biochemistry in the worm has not kept pace. For many reasons, some of which are described in Chapter 1, <italic>C. elegans</italic> is an ideal model to interrogate post-translational modifications that are conserved in higher organisms and those that are conserved in related nematode species. Methods to metabolically label glycoproteins with azide- and alkyne-sugars via endogenous biosynthetic pathways have been described in other species and in some cell lines, but not yet in <italic>C. elegans</italic>. These recent labeling methods importantly enable the downstream purification and identification of specific types of post-translationally modified proteins; however, we were confronted with the lack of available techniques to apply them to <italic>C. elegans</italic>. Thus, in a collaborative effort, we adapted a recently described <italic>C. elegans</italic> primary embryonic cell culture method to the aforementioned labeling strategy to identify glycoproteins (Chapter 2 and Appendix 1). Moreover, we extended this approach to identify <italic>C. elegans</italic> phosphorylcholine modified proteins (Chapter 3). Together, these studies demonstrate that <italic>C. elegans</italic> cells can be utilized in a biochemical approach to study protein post-translational modifications and opens the door to future metabolic labeling experiments. Ultimately, in conjunction with established genetic tools, biochemical characterization of <italic>C. elegans</italic> will make it an even more powerful model organism.