Investigating Glycosylation and Phosphorylcholine Post-Translational Modifications in the Free-Living Nematode, Caenorhabditis elegans

Abstract

Caenorhabditis elegans 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, C. elegans 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 C. elegans. 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 C. elegans. Thus, in a collaborative effort, we adapted a recently described C. elegans 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 C. elegans phosphorylcholine modified proteins (Chapter 3). Together, these studies demonstrate that C. elegans 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 C. elegans will make it an even more powerful model organism.