title: Swainsonine and castanospermine blockade of mannose glycoprotein uptake by macrophages. Apparent inhibition of receptor-mediated endocytosis by endogenous ligands.
abstract: Receptor-mediated uptake of mannose-terminated glycoproteins by macrophages is blocked by treating the cells with swainsonine, an inhibitor of alpha-mannosidase II, and by castanospermine, an inhibitor of the endoplasmic reticulum processing enzyme alpha-glucosidase. Both inhibitors are known to cause accumulation of unprocessed oligosaccharide chains terminating in mannose. Inhibition of ligand uptake by the drugs was time- and dose-dependent. Swainsonine produced a maximal effect after 2 h; castanospermine required 5-6 h. Following swainsonine treatment, complete recovery of mannose receptor activity required 24 h and was blocked by cycloheximide suggesting that new receptor synthesis was necessary. Tunicamycin, an inhibitor of oligosaccharide assembly, had no effect on uptake of mannosylated ligands, but tunicamycin pretreatment reduced the sensitivity to swainsonine. These effects of swainsonine and castanospermine appear to be specific, other macrophage pinocytosis receptors (e.g. mannose phosphate) or phagocytosis of yeast particles were unaffected. Moreover, swainsonine had no effect on the fibroblast mannose phosphate receptor. The ability of macrophages to process newly synthesized oligosaccharides was blocked following treatment with swainsonine. Normal processing was fully recovered 24 h after removal of the drug. Mannosidase II was partially inactivated by swainsonine treatment and only a portion was recovered after 24 h. Treatment of macrophages with swainsonine also resulted in an increase in net lysosomal enzyme secretion. Inhibition of mannose-specific receptor-mediated endocytosis in macrophages by swainsonine and castanospermine appears to be due to the formation of mannose-terminated membrane glycoproteins which engage the mannose receptor thereby preventing function. These results suggest a novel mechanism for regulation of receptor-mediated endocytosis.
mesh_terms: D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D002478:Cells, Cultured; D004705:Endocytosis / Q000187:drug effects*; D005966:Glucuronidase / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D007212:Indolizines; D007700:Kinetics; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D008562:Membrane Glycoproteins; D051381:Rats; D017026:Swainsonine; D014415:Tunicamycin / Q000494:pharmacology; D054836:Indolizidines
chemical_list: D000470:Alkaloids; D005966:Glucuronidase; D006023:Glycoproteins; D007212:Indolizines; D008562:Membrane Glycoproteins; D017026:Swainsonine; D014415:Tunicamycin; C025363:mannoproteins; C037806:castanospermine; D054836:Indolizidines