Oxyrrhis marina is a globally distributed heterotrophic dinoflagellate commonly found in marine and saltwater coastlines. Their cell surface displays an array of polysaccharides, both N- and O-linked. These carbohydrates are used by cells for a multitude of functions including communication, adhesion, and regulation. In a previous study in our lab, changes to the O. marina neutral lipid content during starvation was observed, which ultimately led to curiosity of starvation induced changes to the cell surface, in particular N-linked glycans. In this study, O. marina was used as a model to investigate the changes to the cell’s surface N-glycans under satiated and starvation conditions. The O. marina cells in the satiated condition were actively fed with the prey Isochrysis gambana, a common alga found in saltwater. The prey’s surface glycan profiles were also analyzed as a control. In order to label the surface carbohydrates, they first needed to be taken off the cell itself. The N-glycans were liberated via treatment with PNGaseF, purified by cold ethanol precipitation, and labeled with 2-aminobenzamide (2AB) via reductive amination. Derivatized glycan profiles were analyzed by MALDI-TOF mass spectrometry. The resulting glycan profiles were then annotated using available databases (ie Glycomod) and via manual analysis. PNGaseF effectively liberated N-linked oligosaccharides from glycoproteins from the surface of intact cells. Carbohydrates ionize poorly in mass spectrometry, labeling with 2AB provided a fluorescent label and ionizable group on the glycans through reductive amination. During active feeding on the algae Isochrysis, the O. marina surface should present a specific set of N-linked glycans. No detectable Isochrysis N-glycans were observed in the satiated O. marina. Satiated O. marina displayed a diverse set of high-mannose and hybrid structures. Over a 3-week starvation period, a shift in the O. marina surface N-glycan profile was evident. Under prolonged starvation conditions, the O. marina surface presented a profile composed of truncated glycans modified with phosphate and amino sugars. Our characterization of the O. marina surface provides opportunities for a greater understanding of cell signaling and communication in saturation and starved conditions. In addition, this work provides a first report of the N-linked glycome in this extensively studied marine dinoflagellate.

Investigating changes to the marine dinoflagellate Oxyrrhis marina surface N-glycans during prolonged starvation

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.