Sorting of endocytosed transferrin and asialoglycoprotein occurs immediately after internalization in HepG2 cells

doi:10.1083/jcb.104.5.1261

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Sorting of endocytosed transferrin and asialoglycoprotein occurs immediately after internalization in HepG2 cells

W Stoorvogel, HJ Geuze and GJ Strous

After receptor-mediated uptake, asialoglycoproteins are routed to lysosomes, while transferrin is returned to the medium as apotransferrin. This sorting process was analyzed using 3,3'- diaminobenzidine (DAB) cytochemistry, followed by Percoll density gradient cell fractionation. A conjugate of asialoorosomucoid (ASOR) and horseradish peroxidase (HRP) was used as a ligand for the asialoglycoprotein receptor. Cells were incubated at 0 degree C in the presence of both 131I-transferrin and 125I-ASOR/HRP. Endocytosis of prebound 125I-ASOR/HRP and 131I-transferrin was monitored by cell fractionation on Percoll density gradients. Incubation of the cell homogenate in the presence of DAB and H2O2 before cell fractionation gave rise to a density shift of 125I-ASOR/HRP-containing vesicles due to HRP-catalyzed DAB polymerization. An identical change in density for 125I-transferrin and 125I-ASOR/HRP, induced by DAB cytochemistry, is taken as evidence for the concomitant presence of both ligands in the same compartment. At 37 degrees C, sorting of the two ligands occurred with a half-time of approximately 2 min, and was nearly completed within 10 min. The 125I-ASOR/HRP-induced shift of 131I-transferrin was completely dependent on the receptor-mediated uptake of 125I-ASOR/HRP in the same compartment. In the presence of a weak base (0.3 mM primaquine), the recycling of transferrin receptors was blocked. The cell surface transferrin receptor population was decreased within 6 min to 15% of its original size. DAB cytochemistry showed that sorting between endocytosed 131I-transferrin and 125I-ASOR/HRP was also blocked in the presence of primaquine. These results indicate that transferrin and asialoglycoprotein are taken up via the same compartments and that segregation of the transferrin-receptor complex and asialoglycoprotein occurs very efficiently soon after uptake.
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Olson, J. K., Grose, C. (1998). Complex Formation Facilitates Endocytosis of the Varicella-Zoster Virus gE:gI Fc Receptor. J. Virol. 72: 1542-1551 [Abstract] [Full Text]
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van Helvoort, A, Stoorvogel, W, van Meer, G, Burger, N. (1997). Sphingomyelin synthase is absent from endosomes. J. Cell Sci. 110: 781-788 [Abstract]
Kornilova, E., Sorkina, T., Beguinot, L., Sorkin, A. (1996). Lysosomal Targeting of Epidermal Growth Factor Receptors via a Kinase-dependent Pathway Is Mediated by the Receptor Carboxyl-terminal Residues 1022-1123. J. Biol. Chem. 271: 30340-30346 [Abstract] [Full Text]
Zwart, D. E., Brewer, C. B., Lazarovits, J., Henis, Y. I., Roth, M. G. (1996). Degradation of Mutant Influenza Virus Hemagglutinins Is Influenced by Cytoplasmic Sequences Independent of Internalization Signals. J. Biol. Chem. 271: 907-917 [Abstract] [Full Text]
Thilo, L, Stroud, E, Haylett, T (1995). Maturation of early endosomes and vesicular traffic to lysosomes in relation to membrane recycling. J. Cell Sci. 108: 1791-1803 [Abstract]
Duprez, V, Smoljanovic, M, Lieb, M, Dautry-Varsat, A (1994). Trafficking of interleukin 2 and transferrin in endosomal fractions of T lymphocytes. J. Cell Sci. 107: 1289-1295 [Abstract]
Kok, J. W., Hoekstra, K., Eskelinen, S., Hoekstra, D. (1992). Recycling pathways of glucosylceramide in BHK cells: distinct involvement of early and late endosomes. J. Cell Sci. 103: 1139-1152 [Abstract]
Grewal, T., Heeren, J., Mewawala, D., Schnitgerhans, T., Wendt, D., Salomon, G., Enrich, C., Beisiegel, U., Jackle, S. (2000). Annexin VI Stimulates Endocytosis and Is Involved in the Trafficking of Low Density Lipoprotein to the Prelysosomal Compartment. J. Biol. Chem. 275: 33806-33813 [Abstract] [Full Text]
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