Fluorescently labeled xylosides offer insight into the biosynthetic pathways of glycosaminoglycans.

ABSTRACT

Five novel xylosides tagged with the fluorescent probe Pacific Blue™ were synthesized and found to act as substrates for ß4GalT7, a bottleneck enzyme in the biosynthetic pathways leading to glycosaminoglycans. By confocal microscopy of A549 cells, we showed that the xylosides were taken up by the cells, but did not enter the Golgi apparatus where most of the glycosaminoglycan biosynthesis occurs. Instead, after a possible double galactosylation by ß4GalT7 and ß3GalT6, the biosynthesis was terminated. We hypothesize this is due to the charge of the fluorescent probe, which is required for fluorescent ability and stability under physiological conditions.