Transport of N-acetyl-D-glucosamine and N-acetyl-D-galactosamine by rat liver lysosomes.
Transport of N-acetyl-D-glucosamine and N-acetyl-D-galactosamine, products of lysosomal glycoprotein and glycosaminoglycan degradation, was examined in Percoll gradient purified rat liver lysosomes. Uptake of these two sugars was competitive and quite specific remaining largely unaffected by the presence of L-fucose, D-glucosamine, D-glucose, D-glucuronic acid, D-mannose, or N-acetylneuraminic acid. Kinetic studies revealed a Km of 4.4 mM for both N-acetyl-D-glucosamine and N-acetyl-D-galactosamine uptake. Temperature dependence studies revealed a Q10 of 2.3. N-Acetyl-D-glucosamine uptake was not dependent upon NaCl, KCl, MgCl2, or ATP/MgCl2 and was unaffected by 5 mM dithiothreitol or variation of buffer pH between 6.0 and 8.0. Cytochalasin B at a concentration of 50 microM effectively inhibited uptake of N-acetyl-D-glucosamine by 90% and N-acetyl-D-galactosamine by 65%. Prior incubation of lysosomes in 20 mM N-acetyl-D-glucosamine stimulated uptake of both sugars 3-4-fold, although it had no effect on the uptake of D-glucose. Countertransport was unaffected by neutral and cationic amino acids demonstrating independence from these amino acid transport systems. We conclude that lysosomes possess a highly specific transport system for N-acetyl-D-glucosamine and N-acetyl-D-galactosamine.
Jonas
A J
AJ
Department of Pediatrics, University of Texas Medical School, Houston 77030.
Speller
R J
RJ
Conrad
P B
PB
Dubinsky
W P
WP
D000116:Acetylgalactosamine / Q000378:metabolism*; D000117:Acetylglucosamine / Q000378:metabolism*; D000255:Adenosine Triphosphate; D000818:Animals; D001667:Binding, Competitive; D001692:Biological Transport / Q000187:drug effects; D002412:Cations; D003571:Cytochalasin B; D004229:Dithiothreitol; D005260:Female; D005688:Galactosamine / Q000031:analogs & derivatives*; D005944:Glucosamine / Q000031:analogs & derivatives*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D002412:Cations; D003571:Cytochalasin B; D005688:Galactosamine; D000255:Adenosine Triphosphate; D000116:Acetylgalactosamine; D005944:Glucosamine; D004229:Dithiothreitol; D000117:Acetylglucosamine