Endocytosis and recycling of subunit H1 of the asialoglycoprotein receptor is independent of oligomerization with H2.
The human asialoglycoprotein receptor is composed of two homologous subunits, H1 and H2. By expressing the two subunits in transfected fibroblast cell lines, it has been shown previously that the formation of a hetero-oligomeric complex is necessary for the transport of H2 to the plasma membrane and for high-affinity ligand binding. Here we show that subunit H1, when expressed in the absence of H2, is capable of internalization through coated pits and recycling. The kinetics of these processes are very similar to those of the H1-H2 complex. To study endocytosis in the absence of ligand binding, the cell surface was labeled at 4 degrees C with the 125I-iodinated impermeant reagent sulfosuccinimidyl-3-(4-hydroxyphenyl) propionate, the cells were incubated at 37 degrees C for different times and the amount of internalized receptor was determined by protease digestion of surface proteins and immunoprecipitation. Similarly, recycling of surface-labeled and then internalized receptor protein was studied by monitoring its reappearance on the surface in the presence of exogenous protease. Our results show that subunit H1 contains all the signals necessary for receptor endocytosis and recycling independent of ligand binding.
Geffen
I
I
Department of Biochemistry, University of Basel, Switzerland.
Wessels
H P
HP
Roth
J
J
Shia
M A
MA
Spiess
M
M
D037263:Asialoglycoprotein Receptor; D001692:Biological Transport; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D005347:Fibroblasts; D005786:Gene Expression Regulation; D006207:Half-Life; D006801:Humans; D007150:Immunohistochemistry; D010447:Peptide Hydrolases / Q000378:metabolism; D011971:Receptors, Immunologic / Q000235:genetics / Q000378:metabolism*
D037263:Asialoglycoprotein Receptor; D011971:Receptors, Immunologic; D010447:Peptide Hydrolases