Characterization of a 36,000-dalton protein from the surface of Madin-Darby canine kidney cells involved in cell attachment and spreading.
We have identified and immunochemically characterized a 36,000-dalton membrane glycoprotein from Madin-Darby canine kidney cells. This protein is surface-labeled by lactoperoxidase-mediated iodination and metabolically labeled by [35S]methionine. It binds to Concanavalin A and incorporates 2-D-3H-mannose residues, thus indicating it is a glycoprotein. Rabbit polyclonal antibodies against this protein evenly decorate the external surface of trypsinized, unpolarized cells. The external apical surface of confluent monolayers, grown under culture conditions in which the tight junctions are closed and the cells have acquired polarity, is also evenly stained. The basolateral aspects of the external surface are stained only when the tight junctions are opened by removal of Ca++ or when the antibody has access to the monolayer from the basal side, which indicates an even distribution of this antigen on the surface of polarized cells. The antibody has no inhibitory effect on the opening and resealing of tight junctions in dense cultures, but does inhibit the attachment and spreading of cells on a substrate, which then blocks the establishment of a confluent functional monolayer.
D000818:Animals; D000906:Antibodies / Q000502:physiology; D002448:Cell Adhesion; D002460:Cell Line; D003208:Concanavalin A / Q000378:metabolism; D004285:Dogs; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology / Q000502:physiology; D006651:Histocytochemistry; D007124:Immunoenzyme Techniques; D007163:Immunosorbent Techniques; D007365:Intercellular Junctions / Q000502:physiology; D007668:Kidney / Q000032:analysis* / Q000166:cytology; D008358:Mannose / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis*; D008970:Molecular Weight
D000906:Antibodies; D006023:Glycoproteins; D008565:Membrane Proteins; D003208:Concanavalin A; D008358:Mannose