Monoclonal antibodies identify a group of nuclear pore complex glycoproteins.
Using monoclonal antibodies we identified a group of eight polypeptides of rat liver nuclear envelopes that have common epitopes. Most or all of these proteins are structurally distinct, as shown by tryptic peptide mapping and analysis with polyclonal antibodies. While these polypeptides are relatively tightly bound to nuclear membranes, only one is an integral membrane protein. The eight antigens cofractionate with the nuclear pore complex under various conditions of ionic strength and detergent. It can be seen by immunofluorescence microscopy that the monoclonal antibodies reacting with these antigens stain the nuclear surface of interphase cells in a finely punctate pattern. When the nuclear envelope is disassembled and subsequently reformed during mitosis, the proteins are reversibly dispersed throughout the cytoplasm in the form of minute foci. By EM immunogold localization on isolated nuclear envelopes, the monoclonal antibodies label exclusively the nuclear pore complex, at both its nucleoplasmic and cytoplasmic margins. Considered together, our biochemical and localization data indicate that the eight nuclear envelope polypeptides are pore complex components. As shown in the accompanying paper (Holt, G. D., C. M. Snow, A. Senior, R. S. Haltiwanger, L. Gerace, and G. W. Hart, J. Cell Biol., 104:1157-1164) these eight polypeptides contain a novel form of glycosylation, O-linked N-acetylglucosamine. The relative abundance and disposition of these O-linked glycoproteins in the pore complex are consistent with their having a role in nucleocytoplasmic transport.
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis*; D000939:Epitopes / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis*; D008099:Liver / Q000032:analysis / Q000166:cytology; D008565:Membrane Proteins / Q000032:analysis*; D008970:Molecular Weight; D009685:Nuclear Envelope / Q000032:analysis / Q000648:ultrastructure*; D010449:Peptide Mapping; D051381:Rats
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D006023:Glycoproteins; D008565:Membrane Proteins