Monoclonal antibodies as probes of epithelial membrane polarization.
Monoclonal antibodies directed against antigens in the apical plasma membrane of the toad kidney epithelial cell line A6 were produced to probe the phenomena that underlie the genesis and maintenance of epithelial polarity. Two of these antibodies, 17D7 and 18C3, were selected for detailed study here. 17D7 is directed against a 23-kD peptide found on both the apical and basolateral surfaces of the A6 epithelium whereas 18C3 recognizes a lipid localized to the apical membrane only. This novel observation of an apically localized epithelial lipid species indicates the existence of a specific sorting and insertion process for this, and perhaps other, epithelial plasma membrane lipids. The antibody-antigen complexes formed by both these monoclonal antibodies are rapidly internalized by the A6 cells, but only the 18C3-antigen complex is recycled to the plasma membrane. In contrast to the apical localization of the free antigen, however, the 18C3-antigen complex is recycled to both the apical and basolateral surface of the epithelium, which indicates that monoclonal antibody binding interferes in some way with the normal sorting process for this apical lipid antigen.
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000378:metabolism; D002451:Cell Compartmentation; D002478:Cells, Cultured; D004705:Endocytosis; D004848:Epithelium / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D005089:Exocytosis; D007668:Kidney; D008563:Membrane Lipids / Q000276:immunology / Q000378:metabolism*; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D008970:Molecular Weight; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D008563:Membrane Lipids; D008565:Membrane Proteins