1 research outputs found
Mapping the Expressed Glycome and Glycosyltransferases of Zebrafish Liver Cells as a Relevant Model System for Glycosylation Studies
The emergence of zebrafish as a model organism for human
diseases was accompanied by the development of cellular model systems
that extended the possibilities for <i>in vitro</i> manipulation
and <i>in vivo</i> studies after cell implantation. The
exploitation of zebrafish cell systems is, however, still hampered
by the lack of genomic and biochemical data. Here, we lay a path toward
the efficient use of ZFL, a zebrafish liver-derived cell system, as
a platform for studying glycosylation. To achieve this, we established
the glycomic profile of ZFL by a combination of mass spectrometry
and NMR. We demonstrated that glycoproteins were substituted by highly
sialylated multiantennary <i>N</i>-glycans, some of them
comprising the unusual zebrafish epitope Galβ1–4Â[Neu5AcÂ(α2,3)]ÂGalβ1–4Â[FucÂ(α1,3)]ÂGlcNAc,
and core 1 multisialylated <i>O</i>-glycans. Similarly,
these analyses established that glycolipids were dominated by sialylated
gangliosides. In parallel, analyzing the expression patterns of all
putative sialyl- and fucosyltransferases, we directly correlated the
identified structures to the set of enzymes involved in ZFL glycome.
Finally, we demonstrated that this cell system was amenable to metabolic
labeling using functionalized monosaccharides that permit <i>in vivo</i> imaging of glycosylation processes. Altogether,
glycomics, genomics, and functional studies established ZFL as a relevant
cellular model for the study of glycosylation