title: Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.
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.
authors.affiliation: Department of Cell Biology and Anatomy, Cornell University Medical College, New York, NY 10021.
authors.forename: A
authors.initials: A
authors.lastname: Le Bivic
authors.forename: F X
authors.initials: FX
authors.lastname: Real
authors.forename: E
authors.initials: E
authors.lastname: Rodriguez-Boulan
mesh_terms: D000230:Adenocarcinoma; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D003110:Colonic Neoplasms; D004848:Epithelium / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000096:biosynthesis*; D008854:Microscopy, Electron; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D008565:Membrane Proteins; D009363:Neoplasm Proteins