Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.
- 1 December 1989
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 86 (23) , 9313-9317
- https://doi.org/10.1073/pnas.86.23.9313
Abstract
We studied the surface delivery pathways followed by newly synthesized plasma membrane proteins in intestinal cells. To this end, we developed an assay and characterized an epithelial cell line (SK-CO-15) derived from human colon adenocarcinoma. Polarized confluent monolayers (2000 omega.cm2), grown on polycarbonate filter chambers, were pulsed with radioactive methionine/cysteine and, at different times of chase, the protein fraction reaching the apical or basolateral surface was recovered by domain-selective biotinylation, immunoprecipitation, and immobilized streptavidin precipitation. Both an apical and a basolateral marker were found to be delivered vectorially to the respective surface, with a sorting efficiency of 50:1 for the basolateral marker and 14:1 for the apical marker.Keywords
This publication has 37 references indexed in Scilit:
- Phase separation of integral membrane proteins in Triton X-114 solution.Published by Elsevier ,2021
- Localization by immunofluorescence and histochemical labeling of aminopeptidase N in relation to its biosynthesis in rabbit and pig enterocytesGastroenterology, 1982
- “Western Blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein AAnalytical Biochemistry, 1981
- Biogenesis of intestinal plasma membrane: Posttranslational route and cleavage of sucrase—isomaltaseProceedings of the National Academy of Sciences, 1979
- Fluorographic detection of radioactivity in polyacrylamide gols with the water-soluble fluor, sodium salicylateAnalytical Biochemistry, 1979
- Synthesis of membrane glycoproteins in rat small-intestinal villus cells. Redistribution of l-[1,5,6-3H]-fucose-labelled membrane glycoproteins among Golgi, lateral basal and microvillus membranes in vivoBiochemical Journal, 1979
- Transepithelial transport in cell culture.Proceedings of the National Academy of Sciences, 1976
- Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus.1974
- A TECHNIQUE FOR ULTRACRYOTOMY OF CELL SUSPENSIONS AND TISSUESThe Journal of cell biology, 1973
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970