Benzyl-N-acetyl-alpha-D-galactosaminide induces a storage disease-like phenotype by perturbing the endocytic pathway.
The sugar analog O-benzyl-N-acetyl-alpha-d-galactosaminide (BG) is an inhibitor of glycan chain elongation and inhibits alpha2,3-sialylation in mucus-secreting HT-29 cells. Long-term exposure of these cells to BG is associated with the accumulation of apical glycoproteins in cytoplasmic vesicles. The mechanisms involved therein and the nature of the vesicles have not been elucidated. In these cells, a massive amount of BG metabolites is synthesized. Because sialic acid is mainly distributed apically in epithelial cells, it has been proposed that the BG-induced undersialylation of apical membrane glycoproteins is responsible for their intracellular accumulation due to a defect in anterograde traffic and that sialic acid may constitute an apical targeting signal. In this work, we demonstrate that the intracellular accumulation of membrane glycoproteins does not result mainly from defects in anterograde traffic. By contrast, in BG-treated cells, endocytosed membrane proteins were retained intracellularly for longer periods of time than in control cells and colocalized with accumulated MUC1 and beta(1) integrin in Rab7/lysobisphosphatidic acid(+) vesicles displaying features of late endosomes. The phenotype of BG-treated cells is reminiscent of that observed in lysosomal storage disorders. Sucrose induced a BG-like, lysosomal storage disease-like phenotype without affecting sialylation, indicating that undersialylation is not a requisite for the intracellular accumulation of membrane glycoproteins. Our findings strongly support the notion that the effects observed in BG-treated cells result from the accumulation of BG-derived metabolites and from defects in the endosomal pathway. We propose that abnormal subcellular distribution of membrane glycoproteins involved in cellular communication and/or signaling may also take place in lysosomal storage disorders and may contribute to their pathogenesis.
Ulloa
Fausto
F
Unitat de Biologia Celular i Molecular, Institut Municipal d'Investigació Mèdica, Universitat Pompeu Fabra, Barcelona 08003, Spain.
Real
Francisco X
FX
D000116:Acetylgalactosamine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D001593:Benzyl Compounds / Q000494:pharmacology*; D022162:Cytoplasmic Vesicles / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D019073:HT29 Cells; D006801:Humans; D066298:In Vitro Techniques; D019012:Integrin beta1 / Q000032:analysis; D016464:Lysosomal Storage Diseases / Q000139:chemically induced* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D018396:Mucin-1 / Q000032:analysis; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D010641:Phenotype; D021381:Protein Transport / Q000187:drug effects; D029461:Sialic Acid Storage Disease / Q000139:chemically induced / Q000378:metabolism; D022161:Transport Vesicles / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism
D001593:Benzyl Compounds; D019012:Integrin beta1; D008562:Membrane Glycoproteins; D018396:Mucin-1; C080739:benzyl-alpha-N-acetylgalactosamine; D019158:N-Acetylneuraminic Acid; D000116:Acetylgalactosamine