Receptor-mediated endocytosis of alpha 2-macroglobulin and transferrin in rat caput epididymal epithelial cells in vitro.
The endocytic activity of epithelial cells from the rat epididymis in vitro has been examined by following the uptake of tracer compounds conjugated to proteins. Transferrin-gold and alpha 2-macroglobulin-gold were taken up initially in coated pits, internalized and sequestered into tubular-vesicular structures, multivesicular bodies and, in the case of alpha 2-macroglobulin, into lysosomes. Uptake could be prevented by an excess of unlabeled protein. Studies using 125I-alpha 2-macroglobulin and 125I-transferrin also showed that the uptake of these proteins was specific and could be displaced with increasing amounts of unlabeled protein. In addition, binding of 125I-transferrin to cells was saturable at 4 degrees C. These studies indicate that transferrin and alpha 2-macroglobulin are taken up by receptor-mediated endocytosis. In contrast, a fluid phase marker, bovine serum albumin-gold (BSA-gold), was initially taken up predominantly in uncoated caveolae rather than coated pits, and could not be displaced with excess BSA. By virtue of their charge, polycationized ferritin and unlabeled colloidal gold were taken up and internalized by adsorptive endocytosis, a pathway which is similar to fluid phase endocytosis. The uptake and internalization of alpha 2-macroglobulin and transferrin differed in a number of respects. Uptake and internalization of alpha 2-macroglobulin but not of transferrin was dependent on extracellular calcium. Only alpha 2-macroglobulin was transferred into lysosomes, whereas transferrin was recycled to the cell surface. Although the proton ionophore, monensin, and the transglutaminase inhibitor, dansylcadaverine, did not stop uptake and internalization of either alpha 2-macroglobulin or transferrin, they did prevent the transfer of alpha 2-macroglobulin to lysosomes.
D000327:Adsorption; D000818:Animals; D002478:Cells, Cultured; D003034:Coated Pits, Cell-Membrane / Q000502:physiology; D003102:Colloids; D004705:Endocytosis; D004822:Epididymis / Q000502:physiology* / Q000648:ultrastructure; D004848:Epithelium / Q000502:physiology; D005293:Ferritins / Q000378:metabolism; D006046:Gold; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008297:Male; D008854:Microscopy, Electron; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000502:physiology*; D011971:Receptors, Immunologic / Q000502:physiology*; D011990:Receptors, Transferrin; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism*; D000511:alpha-Macroglobulins / Q000378:metabolism*
D003102:Colloids; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D011990:Receptors, Transferrin; D014168:Transferrin; D000511:alpha-Macroglobulins; D012710:Serum Albumin, Bovine; D006046:Gold; D005293:Ferritins