Differential cell surface expression of four lysosomal integral membrane proteins (LIMPs) in normal rat kidney cells.
Four monoclonal antibodies generated against rat, hepatocyte lysosomal integral membrane protein (LIMPs) (Barriocanal et al., 1986a, b) were used as probes to ascertain the distribution of similar proteins in normal rat kidney (NRK) cells. Comparison of immunoprecipitations of LIMPs 1-4 from hepatocytes and NRK cells revealed a marked similarity in the proteins, in both cell types, as determined by SDS-PAGE. Further, the LIMP epitopes recognized by the antibodies are situated intravesicularly. Ultrastructural immunocytochemistry, using pre-embedding peroxidase, revealed that primary and secondary lysosomes in NRK cells are readily stained with all four antibodies, as well as vesicles in the Golgi region. Immunofluorescence microscopy of non-permeabilized NRK cells with antibodies recognizing LIMPs 1 and 4 illustrated a limited but significant punctate staining pattern of the cell surface. Ultrastructural immunoperoxidase indicated these sites to be cell surface localized coated pits and vesicles. However, it is known that all LIMPs are expressed on the cell surface, albeit at different concentrations, although the total number of each LIMP per cell, respectively, is approximately the same (Barriocanal et al., 1987). Treatment of NRK cells with the acidotropic agent NH4Cl decreased the cell surface expression of LIMPs 1, 3 and 4, but had no effect on LIMP 2. Further, the relative diminution of the cell surface expression varied among the four LIMPs. These results are interpreted to suggest that not all lysosomes contain the same integral membrane proteins in their vesicle container.
Suárez-Quian
C A
CA
Georgetown University, Department of Anatomy and Cell Biology, School of Medicine, Washington, D.C. 20007.
D000818:Animals; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D002478:Cells, Cultured; D011232:Chemical Precipitation; D007668:Kidney / Q000032:analysis* / Q000166:cytology / Q000648:ultrastructure; D008247:Lysosomes / Q000032:analysis* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000032:analysis*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D051381:Rats
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins