Fucose incorporation and identification of fucosylglycoproteins in synaptosomes.
A synaptosome-enriched fraction from sheep cortex was incubated with L-fucose. The uptake of the sugar into this preparation was dependent on time, temperature, and concentration. A Kmapp of 0.94 mM-L-fucose and a Vmaxapp value of 0.24 nM-L-fucose/mg synaptosomal soluble protein/20 min was determined. After incubation for 10 min at 25 degrees C with L-[3H]fucose, 70% of the radioactive label was found in the soluble fraction. DEAE-cellulose chromatography resulted in the elution of three fucosylprotein peaks which were then characterised by gel filtration and sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE). At least eleven 3H protein-staining bands were identified with M. W. 13,000--115,000. Control experiments involving the incubation of the hexose with heat-treated synaptosomes and myelin, mitochondria, and microsomes indicted that the tritiated material associated with the synaptosomal soluble fraction was not due to nonspecific binding or to the presence of contaminating subcellular material. A 3H glycopeptide was identified, and on analysis the carbohydrate moiety was found to be rich in sialic acid, fucose, galactose, mannose, and N-acetylglucosamine. Mild acid treatment of the glycopeptide released fucose, which implies that this carbohydrate occupies a terminal position in the oligosaccharide chain. From these results it is proposed that synthesis or the modification of soluble fucosylglycoproteins is possible in synaptosomes.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001692:Biological Transport; D002241:Carbohydrates / Q000032:analysis; D002540:Cerebral Cortex / Q000378:metabolism; D005643:Fucose / Q000378:metabolism*; D006020:Glycopeptides / Q000032:analysis; D006023:Glycoproteins / Q000096:biosynthesis*; D007700:Kinetics; D012756:Sheep; D013347:Subcellular Fractions / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism*; D013696:Temperature
D000596:Amino Acids; D002241:Carbohydrates; D006020:Glycopeptides; D006023:Glycoproteins; D005643:Fucose