Purification and characterization of the L-fucose transporter.
L-Fucose is a monosaccharide present in low levels in the serum. It is, however, a common structural component of glycoproteins. L-Fucose is accumulated in eukaryotic cells by a specific, facilitative diffusion transport system which has been designated the fucose transporter. In this study, purification of the transporter from mouse brain was performed by detergent extraction followed by ion-exchange and reactive dye ligand column chromatography. Purification was followed using a transport assay into reconstituted liposomes. A 111-fold purification with 5% yield was achieved from the crude homogenate. The apparent molecular weight of the protein was 57 kDa. Transport was found to be saturable. The K(m) and V(max) values are estimated at 3 microM and 275 pmol/min/mg, respectively. The tissue distribution of fucose transport was examined in liver, kidney, heart, lung, spleen, brain, muscle, adipose, ovary, pancreas, and thymus. Some fucose transport was found in all tissues examined. Very low levels were observed in the liver relative to all other tissues examined. The only monosaccharide which could inhibit the uptake of L-[5,6-(3)H]fucose was fucose itself.
Leck
Joshua R
JR
Department of Chemistry, Fort Hays State University 600 Park Street, Hays, KS 67601, USA.
Wiese
Thomas J
TJ
D000818:Animals; D001692:Biological Transport; D001921:Brain / Q000196:embryology; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005643:Fucose / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry; D007700:Kinetics; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D014018:Tissue Distribution
D003902:Detergents; D006023:Glycoproteins; C499067:L-fucose transporter protein, mouse; D009004:Monosaccharide Transport Proteins; D005643:Fucose